Cxcr antagonistic peptides and uses thereof

ABSTRACT

The present invention provides a novel class of medicament based on CXCR antagonistic peptides. Among other things, the present invention provides peptides, compositions and methods for treating diseases, disorders and conditions in which a CXCR mediated pathway is implicated. Compositions and methods for effective treatment of inflammation, stroke, traumatic brain injury, pancreatic cancer, and others are provided.

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application Ser.No. 61/908,307 filed Nov. 25, 2013, the disclosure of which is herebyincorporated by reference.

BACKGROUND

CXC chemokine receptors are a subfamily of chemokine receptors thatspecifically bind and respond to cytokines of the CXC chemokine family.There are currently seven known CXC chemokine receptors in mammals,named CXCR1 through CXCR7. CXC receptors have been implicated in variousinflammatory, autoimmune and other diseases, disorders or conditions.

Inflammation-related diseases, disorders or conditions include a varietyof specific pathologies affecting millions of people. While severaltherapies have been developed, such as certain non-steroidalanti-inflammatory compounds for use in the treatment of rheumatoidarthritis, there is still a large unmet need for better and moreefficacious therapies.

SUMMARY OF THE INVENTION

The present invention provides a novel class of medicament based on CXCRantagonistic peptides. Among other things, the present inventionprovides peptides, compositions and methods for treating diseases,disorders and conditions in which a CXCR mediated pathway is implicated.Thus, the present invention is particularly useful for effectivetreatment of inflammation, stroke, traumatic brain injury, pancreaticcancer, neurodegenerative diseases, to name but a few.

In one aspect, the present invention provides a peptide of less than 6amino acids comprising the amino acid sequence proline-glycine-X (PGX)for use as a medicament, wherein X is any amino acid.

In another aspect, the present invention provides a peptide of less than6 amino acids comprising the amino acid sequence proline-glycine-X (PGX)for use in the treatment of inflammation, wherein X is any amino acid.In some embodiments, the inflammation is associated with arthritis(including rheumatoid arthritis and juvenile rheumatoid arthritis,);vasculitis; multiple sclerosis; autoimmune thyroiditis; inflammatorybowel disease (IBD); Crohn's disease; inflammatory conditions of thenervous system (e.g., Alzheimer's disease), liver (e.g., hepatitis),kidney (e.g., nephritis) and pancreas (e.g., pancreatitis);cardiovascular disorders (e.g., pulmonary fibrosis, idiopathic pulmonaryfibrosis, reperfusion injury, acute vaso-occlusive crisis in sickle cellanemia); respiratory disorders, e.g., COPD (e.g., cystic fibrosis); andinfectious disease (e.g., acute endocarditis, pericarditis); adultrespiratory distress syndrome (ARDS); Chronic Obstructive Pulmonarydisease (COPD); tumors or cancers (e.g., soft tissue or solid tumors),such as melanoma or prostate cancer; or transplant rejection (e.g.destruction of pancreatic islet cells in islet cell transplantation,delayed graft failure in kidney or other organ transplantation); acuteand chronic graft versus host disease. In some embodiments, theinflammation is associated with an acute inflammatory condition. In someembodiments, the inflammation is associated with a chronic inflammatorycondition. In some embodiments, the inflammation is neuroinflammationassociated with stroke.

In still another aspect, the present invention provides a peptide ofless than 6 amino acids comprising the amino acid sequenceproline-glycine-X (PGX) for use in the treatment of stroke, wherein X isany amino acid.

In another aspect, the present invention provides a peptide of less than6 amino acids comprising the amino acid sequence proline-glycine-X (PGX)for use in the treatment of traumatic brain injury, wherein X is anyamino acid.

In yet another aspect, the present invention provides a peptide of lessthan 6 amino acids comprising the amino acid sequence proline-glycine-X(PGX) for use in the treatment of pancreatic cancer, wherein X is anyamino acid.

In further aspect, the present invention provides a peptide of less than6 amino acids comprising the amino acid sequence proline-glycine-X (PGX)for use in the treatment or prevention of neuronal death, wherein X isany amino acid. In some embodiments, neuronal death is associated with aneurodegenerative disease. In some embodiments, the neurodegenerativedisease is selected from the group consisting of amyotrophic lateralsclerosis (ALS), Huntington's disease, Alzheimer's disease, Parkinson'sdisease, multiple sclerosis, vascular cognitive impairment (VCI), andcombinations thereof.

In another aspect, the present invention provides methods of treatinginflammation, comprising administering to a subject in need thereof apeptide of less than 6 amino acids containing the amino acid sequenceproline-glycine-X (PGX), wherein X is any amino acid, or a compositioncontaining such peptide.

In some embodiments, the inflammation is associated with arthritis(including rheumatoid arthritis and juvenile rheumatoid arthritis,);vasculitis; multiple sclerosis; autoimmune thyroiditis; inflammatorybowel disease (IBD); Crohn's disease; inflammatory conditions of, thenervous system (e.g., Alzheimer's disease), liver (e.g., hepatitis),kidney (e.g., nephritis) and pancreas (e.g., pancreatitis);cardiovascular disorders (pulmonary fibrosis, idiopathic pulmonaryfibrosis, reperfusion injury, acute vaso-occlusive crisis in sickle cellanemia); respiratory disorders, e.g., COPD (e.g., cystic fibrosis); andinfectious disease (e.g., acute endocarditis, pericarditis); adultrespiratory distress syndrome (ARDS); Chronic Obstructive Pulmonarydisease (COPD); tumors or cancers (e.g., soft tissue or solid tumors),such as melanoma or prostate cancer; and/or transplant rejection (e.g.destruction of pancreatic islet cells in islet cell transplantation,delayed graft failure in kidney or other organ transplantation); acuteand chronic graft versus host disease. In some embodiments, theinflammation is associated with an acute inflammatory condition. In someembodiments, the inflammation is associated with a chronic inflammatorycondition. In some embodiments, the inflammation is neuroinflammationassociated with stroke.

In another aspect, the present invention provides methods of treatingstroke, comprising administering to a subject in need of treatment apeptide of less than 6 amino acids containing the amino acid sequencePGX, wherein X is any amino acid, or a composition comprising suchpeptide.

In another aspect, the present invention provides methods of treatingbrain injury (e.g., traumatic brain injury), comprising administering toa subject in need of treatment a peptide of less than 6 amino acidscontaining the amino acid sequence PGX, wherein X is any amino acid, ora composition comprising such peptide.

In another aspect, the present invention provides methods of treatingpancreatic cancer, comprising administering to a subject in need oftreatment a peptide of less than 6 amino acids containing the amino acidsequence PGX, wherein X is any amino acid, or a composition comprisingsuch peptide.

In further aspect, the present invention provides methods of treating orpreventing neuronal death, comprising administering to a subject in needof treatment a peptide of less than 6 amino acids containing the aminoacid sequence of PGX, wherein X is any amino acid, or a compositioncomprising such peptide. In some embodiments, the neuronal death isassociated with a neurodegenerative disease. In some embodiments, theneurodegenerative disease is selected from the group consisting ofamyotrophic lateral sclerosis (ALS), Huntington's disease, Alzheimer'sdisease, Parkinson's disease, multiple sclerosis, vascular cognitiveimpairment (VCI), and combinations thereof.

In another aspect, the present invention provides methods of inhibitinga CXC receptor activity in a cell, comprising administering to a cellpeptides of less than 6 amino acids containing the amino acid sequenceproline-glycine-X (PGX), wherein X is any amino acid. In someembodiments, the cell is a mammalian cell. In some embodiments, the cellis a human cell. In some embodiments, the cell is a cultured cell. Insome embodiments, the cell is in a living organism. In some embodiments,the CXC receptor is selected from CXCR1, CXCR2, CXCR3, CXCR4, CXCR5,CXCR6, and/or CXCR7. In some embodiments, the CXC receptor is CXCR1 orCXCR2.

In still another aspect, the present invention provides a method ofinhibiting a CXC receptor activity in a subject, comprisingadministering to a subject in need thereof a peptide of less than 6amino acids containing the amino acid sequence proline-glycine-X (PGX),wherein X is any amino acid.

In various embodiments, a suitable peptide contains the amino acidsequence proline-glycine-X (PGX), wherein X is any amino acid exceptproline. In various embodiments, a suitable peptide contains the aminoacid sequence proline-glycine-X (PGX), wherein X is glycine. In someembodiments, the amino acid sequence proline-glycine-X (PGX) is at theN-terminus. In some embodiments, a suitable peptide is acetylated ormethylated at the N-terminus. In some embodiments, a suitable peptidehas less than 5 amino acids. In some embodiments, a suitable peptide hasless than 4 amino acids. In some embodiments, a suitable peptide is theN-acetylated tripeptide ac-PGG. In some embodiments, a suitable peptideis PEGylated.

In some embodiments, a suitable peptide contains a plurality of PGXrepeats. In some embodiments, a suitable peptide contains 2-50, 2-40,2-30, 2-20, 2-15, 2-10, 2-8, 2-6, or 2-5 PGX repeats. In someembodiments, a suitable peptide contains more than 2, 3, 4, 5, 10, 15,20, 25, 30, 35, 40, 45 or 50 PGX repeats. In some embodiments, asuitable peptide contains PGX repeats wherein the PGX repeats are tandemrepeats. In some embodiments, a suitable peptide contains PGX repeatswherein the PGX repeats are interrupted by non-PGX sequences. In someembodiments, a suitable peptide contains PGX repeats wherein the PGXrepeats are linear and/or branched.

In various embodiments, a suitable peptide is administeredintravenously, subcutaneously, orally, transdermally, intramuscularly,intraperitoneally, and/or by aerosol inhalation.

In still another aspect, the present invention provides pharmaceuticalcompositions comprising a therapeutically effective amount of a peptide(e.g., of less than 6, 5, or 4 amino acids) and containing a sequence ofPGX, wherein X is any amino acid except proline and a pharmaceuticallyacceptable carrier or excipient. In some embodiments, X is any aminoacid except proline. In some embodiments, X is glycine. In someembodiments, a pharmaceutical composition according to the inventioncontains a peptide that is PEGylated. In some embodiments, apharmaceutical composition according to the invention contains a peptidewherein the sequence of proline-glycine-X (PGX) is at the N-terminus. Insome embodiments, a pharmaceutical composition according to theinvention contains a peptide acetylated or methylated at the N-terminus.In some embodiments, a pharmaceutical composition according to theinvention contains an N-acetylated tripeptide ac-PGG. In someembodiments, a pharmaceutical composition according to the inventioncontains a peptide with a plurality of PGX repeats. In some embodiments,a pharmaceutical composition according to the invention contains apeptide with 2-50, 2-40, 2-30, 2-20, 2-15, 2-10, 2-8, 2-6, or 2-5 PGXrepeats. In some embodiments, a pharmaceutical composition according tothe invention contains a peptide with more than 2, 3, 4, 5, 10, 15, 20,25, 30, 35, 40, 45 or 50 PGX repeats. In some embodiments, apharmaceutical composition according to the invention contains a peptidewith PGX repeats that are tandem repeats. In some embodiments, apharmaceutical composition according to the invention contains a peptidewith PGX repeats wherein the PGX repeats are interrupted by non-PGXsequences. In some embodiments, a pharmaceutical composition accordingto the invention contains a peptide with PGX repeats wherein the PGXrepeats are linear and/or branched.

As used in this application, the terms “about” and “approximately” areused as equivalents. Any numerals used in this application with orwithout about/approximately are meant to cover any normal fluctuationsappreciated by one of ordinary skill in the relevant art.

Other features, objects, and advantages of the present invention areapparent in the detailed description that follows. It should beunderstood, however, that the detailed description, while indicatingembodiments of the present invention, is given by way of illustrationonly, not limitation. Various changes and modifications within the scopeof the invention will become apparent to those skilled in the art fromthe detailed description.

BRIEF DESCRIPTION OF THE DRAWING

The drawings are for illustration purposes only, not for limitation.

FIG. 1 depicts exemplary graphs showing the effects of pretreatment ofneurons with several doses of SB225002, anti-CXCR2 ligand binding domainantibody, or ac-PGG on ac-PGP-induced apoptosis as measured via a TUNELstain. A. Cells pretreated with all doses of SB225002 were significantlyless apoptotic than cells treated with ac-PGP only (*). Cells pretreatedwith 1 nM SB225002 were significantly less apoptotic than cells treatedwith 0.01 and 0.1 nM SB225002 (#). B. Cells pretreated with CXCR2antibody at a 1:1000 (1e3) or greater dilution exhibited significantlyless apoptosis than ac-PGP alone (*) while a 1e3 dilution producedsignificantly less apoptosis than a 1e2 dilution (#). C. Neuronspretreated with all doses of ac-PGG were significantly less apoptoticthan ac-PGP treatment alone (*). Cells pretreated with 100 nM ac-PGGwere significantly less apoptotic than cells pretreated with 0.01 nMac-PGG (#).

FIG. 2 depicts an exemplary graph showing the effects of pretreatmentwith vehicle, SB225002, CXCR2 antibody, ac-PGG or ac-PGP on neuronssubjected to 2-hour oxygen-glucose deprivation (OGD). Significantapoptosis was induced in vehicle- and ac-PGP treated neurons compared tonormoxic controls (*). Neurons pretreated with SB225002, CXCR2 antibody,or ac-PGG were significantly less apoptotic than vehicle-treatedcontrols (#).

FIG. 3 depicts an exemplary graph showing the effects of pretreatment ofneurons with PGG on ac-PGP-induced apoptosis as measured via TUNELstain. Neurons pretreated with all doses of PGG were significantly lessapoptotic than ac-PGP treatment alone (*). Cells pretreated with 100 nMPGG were significantly less apoptotic than cells pretreated with 0.01 nMPGG (#).

FIG. 4 depicts exemplary graphs showing the effects of treatment withvehicle, SB225002, CXCR2 antibody, or ac-PGG on neurons at reoxygenationor 5, 15, 30, or 60 minutes after reoxygenation in an OGD paradigm. A.Treatment with SB225002 resulted in significant protection from OGD atall time points compared to vehicle (*). When administered at 15 minutespost-reoxygenation, neuronal protection by SB225002 significantlydecreased compared to administration at reoxygenation (#). B. Treatmentwith CXCR2 antibody resulted in significant protection from OGD at alltime points compared to vehicle (*). When administered at 60 minutespost-reoxygenation, neuronal protection by CXCR2 antibody significantlydecreased compared to administration at reoxygenation (#). C. Treatmentwith ac-PGG resulted in significant protection from OGD at all timepoints compared to vehicle (*). No change in the efficacy of neuronalprotection by ac-PGG was observed when administered up to 60 minutesafter reoxygenation.

FIG. 5 depicts an exemplary graph showing the effects of treatment with1, 10 and 100 mg/kg doses of ac-PGG on total infarct volume in a strokemodel with Spontaneously Hypertensive Rats (SHR). Treatment with ac-PGGdecreased total infarct volume when administered at reperfusion.

FIG. 6 depicts an exemplary graph showing the effects of treatment with1, 10 and 100 mg/kg doses of ac-PGG on percent reduction in infarctvolume in a stroke model with Spontaneously Hypertensive Rats (SHR).Treatment with ac-PGG increased percent reduction in infarct volume whenadministered at reperfusion.

FIG. 7 depicts representative TCC-stained brain sections showingdifferences in infarction between animals receiving vehicle or 100 mg/kgac-PGG.

FIG. 8 depicts an exemplary graph showing the effects of 0.1, 10, 1, and50 mg/kg doses of ac-PGG on a Carrageenan-induced change in paw volume.The 3 highest doses of ac-PGG decreased the Carrageenan-induced increasein paw volume at 3, 4, and 5 hours post-Carrageenan injection.

FIG. 9 depicts an exemplary graph showing the effects of 0.1, 10, 1, and50 mg/kg doses of ac-PGG on latency to respond to a hot plate. The 1mg/kg and 50 mg/kg doses of ac-PGG decreased change in latency to hotplate response 3 hours after Carrageenan injection, while only the 50mg/kg dose lead to a significant decrease in latency change 5 hoursafter Carrageenan injection.

FIG. 10 depicts an exemplary graph showing the effects of two dosingschedules of ac-PGG on EAE score. Both ac-PGG dosing schedules decreasedEAE scores compared to the Vehicle group.

FIG. 11 depicts an exemplary graph showing the effects of two dosingschedules of ac-PGG on weight. Both ac-PGG dosing schedules increasedweight compared to the Vehicle group over the course of the study.

FIG. 12 depicts exemplary MRI pictures from Day 14 of EAE inductionshowing the effects of treatment with ac-PGG (days 8-15) on white matterhyperintensities. Treatment with ac-PGG decreased the number of whitematter hyperintensities seen with T2-weighted MRI scans.

DEFINITIONS

In order for the present invention to be more readily understood,certain terms are first defined below. Additional definitions for thefollowing terms and other terms are set forth throughout thespecification.

Agonist: As used herein, the term “agonist” refers to any molecule thathas a positive impact in a function of a protein of interest. In someembodiments, an agonist directly or indirectly enhances, strengthens,and/or increases an activity of a protein of interest. In particularembodiments, an agonist directly interacts with the protein of interest.Agonists can be, e.g., proteins, chemical compounds, small molecules,nucleic acids, antibodies, drugs, ligands, or other agents.

Amino acid: As used herein, term “amino acid,” in its broadest sense,refers to any compound and/or substance that can be incorporated into apolypeptide chain. In some embodiments, an amino acid has the generalstructure H2N—C(H)(R)—COOH. In some embodiments, an amino acid is anaturally occurring amino acid. In some embodiments, an amino acid is asynthetic amino acid; in some embodiments, an amino acid is a D-aminoacid; in some embodiments, an amino acid is an L-amino acid. “Standardamino acid” refers to any of the twenty standard L-amino acids commonlyfound in naturally occurring peptides. “Nonstandard amino acid” refersto any amino acid, other than the standard amino acids, regardless ofwhether it is prepared synthetically or obtained from a natural source.As used herein, “synthetic amino acid” encompasses chemically modifiedamino acids, including but not limited to salts, amino acid derivatives(such as amides), and/or substitutions. Amino acids, including carboxy-and/or amino-terminal amino acids in peptides, can be modified b_(y)methylation, amidation, acetylation, protecting groups, and/orsubstitution with other chemical groups that can change the peptide'scirculating half-life without adversely affecting their activity.Examples of unconventional or un-natural amino acids include, but arenot limited to, citrulline, ornithine, norleucine, norvaline,4-(E)-butenyl-4(R)-methyl-N-methylthreonine (MeBmt), N-methyl-leucine(MeLeu), aminoisobutyric acid, statin, and N-methyl-alanine (MeAla).Amino acids may participate in a disulfide bond. Amino acids maycomprise one or posttranslational modifications, such as associationwith one or more chemical entities (e.g., methyl groups, acetate groups,acetyl groups, phosphate groups, formyl moieties, isoprenoid groups,sulfate groups, polyethylene glycol moieties, lipid moieties,carbohydrate moieties, biotin moieties, etc. The term “amino acid” isused interchangeably with “amino acid residue,” and may refer to a freeamino acid and/or to an amino acid residue of a peptide. It will beapparent from the context in which the term is used whether it refers toa free amino acid or a residue of a peptide.

Animal: As used herein, the term “animal” refers to any member of theanimal kingdom. In some embodiments, “animal” refers to humans, at anystage of development. In some embodiments, “animal” refers to non-humananimals, at any stage of development. In certain embodiments, thenon-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit,a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig). Insome embodiments, animals include, but are not limited to, mammals,birds, reptiles, amphibians, fish, insects, and/or worms. In someembodiments, an animal may be a transgenic animal,genetically-engineered animal, and/or a clone.

Antagonist: As used herein, the term “antagonist” refers to any moleculethat has a negative impact in a function of a protein of interest. Insome embodiments, an antagonist directly or indirectly reduces,inhibits, or down-regulates an activity of a protein of interest. Inparticular embodiments, an antagonist directly interacts with theprotein of interest. Antagonists can be, e.g., proteins, chemicalcompounds, small molecules, nucleic acids, antibodies, drugs, ligands,or other agents.

Approximately or about: As used herein, the term “approximately” or“about,” as applied to one or more values of interest, refers to a valuethat is similar to a stated reference value. In certain embodiments, theterm “approximately” or “about” refers to a range of values that fallwithin 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%,8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greaterthan or less than) of the stated reference value unless otherwise statedor otherwise evident from the context (except where such number wouldexceed 100% of a possible value).

Carrier or diluent: As used herein, the terms “carrier” and “diluent”refers to a pharmaceutically acceptable (e.g., safe and non-toxic foradministration to a human) carrier or diluting substance useful for thepreparation of a pharmaceutical formulation. Exemplary diluents includesterile water, bacteriostatic water for injection (BWFI), a pH bufferedsolution (e.g. phosphate-buffered saline), sterile saline solution,Ringer's solution or dextrose solution.

Dosage form: As used herein, the terms “dosage form” and “unit dosageform” refer to a physically discrete unit of a therapeutic agent for thepatient to be treated. Each unit contains a predetermined quantity ofactive material calculated to produce the desired therapeutic effect. Itwill be understood, however, that the total dosage of the compositionwill be decided by the attending physician within the scope of soundmedical judgment.

Dosing regimen: A “dosing regimen” (or “therapeutic regimen”), as thatterm is used herein, is a set of unit doses (typically more than one)that are administered individually to a subject, typically separated byperiods of time. In some embodiments, a given therapeutic agent has arecommended dosing regimen, which may involve one or more doses. In someembodiments, a dosing regimen comprises a plurality of doses each ofwhich are separated from one another by a time period of the samelength; in some embodiments, a dosing regimen comprises a plurality ofdoses and at least two different time periods separating individualdoses. In some embodiments, the therapeutic agent is administeredcontinuously over a predetermined period. In some embodiments, thetherapeutic agent is administered once a day (QD) or twice a day (BID).

Functional equivalent or derivative: As used herein, the term“functional equivalent” or “functional derivative” denotes a moleculethat retains a biological activity (either function or structural) thatis substantially similar to that of the original sequence. A functionalderivative or equivalent may be a natural derivative or is preparedsynthetically. Exemplary functional derivatives include amino acidsequences having substitutions, deletions, or additions of one or moreamino acids, provided that the biological activity of the protein isconserved. The substituting amino acid desirably has chemico-physicalproperties which are similar to that of the substituted amino acid.Desirable similar chemico-physical properties include, similarities incharge, bulkiness, hydrophobicity, hydrophilicity, and the like.

Improve, increase, or reduce: As used herein, the terms “improve,”“increase” or “reduce,” or grammatical equivalents, indicate values thatare relative to a baseline measurement, such as a measurement in thesame individual prior to initiation of the treatment described herein,or a measurement in a control individual (or multiple controlindividuals) in the absence of the treatment described herein. A“control individual” is an individual afflicted with the same form ofdisease as the individual being treated, who is about the same age asthe individual being treated (to ensure that the stages of the diseasein the treated individual and the control individual(s) are comparable).

In vitro: As used herein, the term “in vitro” refers to events thatoccur in an artificial environment, e.g., in a test tube or reactionvessel, in cell culture, etc., rather than within a multi-cellularorganism.

In vivo: As used herein, the term “in vivo” refers to events that occurwithin a multi-cellular organism, such as a human and a non-humananimal. In the context of cell-based systems, the term may be used torefer to events that occur within a living cell (as opposed to, forexample, in vitro systems).

Prevent: As used herein, the term “prevent” or “prevention”, when usedin connection with the occurrence of a disease, disorder, and/orcondition, refers to reducing the risk of developing the disease,disorder and/or condition. See the definition of “risk.”

Polypeptide: The term “polypeptide” as used herein refers to asequential chain of amino acids linked together via peptide bonds. Theterm is used to refer to an amino acid chain of any length, but one ofordinary skill in the art will understand that the term is not limitedto lengthy chains and can refer to a minimal chain comprising two aminoacids linked together via a peptide bond. As is known to those skilledin the art, polypeptides may be processed and/or modified. The terms“peptide” and “polypeptide” are used interchangeably herein.

Protein: The term “protein” as used herein refers to one or morepolypeptides that function as a discrete unit. If a single polypeptideis the discrete functioning unit and does not require permanent ortemporary physical association with other polypeptides in order to formthe discrete functioning unit, the terms “polypeptide” and “protein” maybe used interchangeably. If the discrete functional unit is comprised ofmore than one polypeptide that physically associate with one another,the term “protein” refers to the multiple polypeptides that arephysically coupled and function together as the discrete unit.

Risk: As will be understood from context, a “risk” of a disease,disorder, and/or condition comprises a likelihood that a particularindividual will develop a disease, disorder, and/or condition (e.g., aninflammation-related disease, disorder, or condition). In someembodiments, risk is expressed as a percentage. In some embodiments,risk is from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70,80, 90 up to 100%. In some embodiments risk is expressed as a riskrelative to a risk associated with a reference sample or group ofreference samples. In some embodiments, a reference sample or group ofreference samples have a known risk of a disease, disorder, conditionand/or event (e.g., an inflammation-related disease, disorder, orcondition). In some embodiments a reference sample or group of referencesamples are from individuals comparable to a particular individual. Insome embodiments, relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, ormore.

Stability: As used herein, the term “stable” refers to the ability ofthe therapeutic agent to maintain its therapeutic efficacy (e.g., all orthe majority of its intended biological activity and/or physiochemicalintegrity) over extended periods of time. The stability of a therapeuticagent, and the capability of the pharmaceutical composition to maintainstability of such therapeutic agent, may be assessed over extendedperiods of time (e.g., for at least 1, 3, 6, 12, 18, 24, 30, 36 monthsor more). In certain embodiments, pharmaceutical compositions describedherein have been formulated such that they are capable of stabilizing,or alternatively slowing or preventing the degradation, of one or moretherapeutic agents formulated therewith. In the context of a formulationa stable formulation is one in which the therapeutic agent thereinessentially retains its physical and/or chemical integrity andbiological activity upon storage and during processes (such asfreeze/thaw, mechanical mixing and lyophilization).

Subject: As used herein, the term “subject” refers to a human or anynon-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine,sheep, horse or primate). A human includes pre and post natal forms. Inmany embodiments, a subject is a human being. A subject can be apatient, which refers to a human presenting to a medical provider fordiagnosis or treatment of a disease. The term “subject” is used hereininterchangeably with “individual” or “patient.” A subject can beafflicted with or is susceptible to a disease or disorder but may or maynot display symptoms of the disease or disorder.

Substantially: As used herein, the term “substantially” refers to thequalitative condition of exhibiting total or near-total extent or degreeof a characteristic or property of interest. One of ordinary skill inthe biological arts will understand that biological and chemicalphenomena rarely, if ever, go to completion and/or proceed tocompleteness or achieve or avoid an absolute result. The term“substantially” is therefore used herein to capture the potential lackof completeness inherent in many biological and chemical phenomena.

Suffering from: An individual who is “suffering from” a disease,disorder, and/or condition has been diagnosed with or displays one ormore symptoms of the disease, disorder, and/or condition.

Susceptible to: An individual who is “susceptible to” a disease,disorder, and/or condition has not been diagnosed with the disease,disorder, and/or condition. In some embodiments, an individual who issusceptible to a disease, disorder, and/or condition may not exhibitsymptoms of the disease, disorder, and/or condition. In someembodiments, an individual who is susceptible to a disease, disorder,condition, or event may be characterized by one or more of thefollowing: (1) a genetic mutation associated with development of thedisease, disorder, and/or condition; (2) a genetic polymorphismassociated with development of the disease, disorder, and/or condition;(3) increased and/or decreased expression and/or activity of a proteinassociated with the disease, disorder, and/or condition; (4) habitsand/or lifestyles associated with development of the disease, disorder,condition, and/or event (5) having undergone, planning to undergo, orrequiring a transplant. In some embodiments, an individual who issusceptible to a disease, disorder, and/or condition will develop thedisease, disorder, and/or condition. In some embodiments, an individualwho is susceptible to a disease, disorder, and/or condition will notdevelop the disease, disorder, and/or condition.

Therapeutically effective amount: As used herein, the term“therapeutically effective amount” of a therapeutic agent means anamount that is sufficient, when administered to a subject suffering fromor susceptible to a disease, disorder, and/or condition, to treat,diagnose, prevent, and/or delay the onset of the symptom(s) of thedisease, disorder, and/or condition. It will be appreciated by those ofordinary skill in the art that a therapeutically effective amount istypically administered via a dosing regimen comprising at least one unitdose.

Treating: As used herein, the term “treat,” “treatment,” or “treating”refers to any method used to partially or completely alleviate,ameliorate, relieve, inhibit, prevent, delay onset of, reduce severityof and/or reduce incidence of one or more symptoms or features of aparticular disease, disorder, and/or condition. Treatment may beadministered to a subject who does not exhibit signs of a disease and/orexhibits only early signs of the disease for the purpose of decreasingthe risk of developing pathology associated with the disease.

DETAILED DESCRIPTION

The present invention provides, among other things, a novel class ofmedicaments based on CXCR antagonistic peptides. In particular, thepresent invention provides compositions and methods for treatment ofCXCR related diseases, disorders and conditions.

Various aspects of the invention are described in detail in thefollowing sections. The use of sections is not meant to limit theinvention. Each section can apply to any aspect of the invention. Inthis application, the use of “or” means “and/or” unless statedotherwise.

CXC Receptors

Generally speaking, CXC receptors (CXCR) are chemokine receptors thatbind and respond to cytokines of the CXC chemokine family. Typically,they are members of G protein-linked receptors, also known as seventransmembrane (7-TM) proteins, because they are characterized withspanning the cell membrane seven times. CXCRs according to the presentinvention include, but not limited to, CXCR1 through CXCR7 (i.e., CXCR1,CXCR2, CXCR3, CXCR4, CXCR5, CXCR6 and CXCR7).

CXCR1 and CXCR2 are closely related receptors that recognize CXCchemokines that possess an E-L-R amino acid motif immediately adjacentto their CXC motif. CXCL8 (otherwise known as interleukin-8) and CXCL6can both bind CXCR1 in humans, while all other ELR-positive chemokines,such as CXCL1 to CXCL7 bind only CXCR2. They are both expressed on thesurface of neutrophils in mammals.

CXCR3 is expressed predominantly on T lymphocytes, and also on otherlymphocytes (some B cells and NK cells) and is highly induced followingcell activation. There are two isoforms, CXCR3-A and CXCR3-B. It hasthree highly related ligands in mammals, CXCL9, CXCL10 and CXCL11.

CXCR4 (also known as fusin) is the receptor for a chemokine known asCXCL12 (or SDF-1) and, as with CCR5, is utilized by HIV-1 to gain entryinto target cells. This receptor has a wide cellular distribution, withexpression on most immature and mature hematopoietic cell types (e.g.,neutrophils, monocytes, T and B cells, dendritic cells, Langerhans cellsand macrophages). In addition, CXCR4 can also be found on vascularendothelial cells and neuronal/nerve cells.

CXCR5 is selectively expressed on B cells and is involved in lymphocytehoming and the development of normal lymphoid tissue. Its principleligand is CXCL13 (or BLC).

CXCR6 was formerly called three different names (STRL33, BONZO, andTYMSTR) before being assigned CXCR6 based on its chromosomal location(within the chemokine receptor cluster on human chromosome 3p21) and itssimilarity to other chemokine receptors in its gene sequence. CXCR6binds the ligand CXCL16. However, CXCR6 is more closely related instructure to CC chemokine receptors than to other CXC chemokinereceptors.

CXCR7 was originally called RDC-1 (an orphan receptor) but has sincebeen shown to cause chemotaxis in T lymphocytes in response to CXCL12(the ligand for CXCR4) prompting the renaming of this molecule as CXCR7.There is no information publicly available to confirm whether thisdesignation has been accepted by the IUIS/WHO Subcommittee on ChemokineNomenclature at this time. This receptor has also been identified onmemory B cells.

CXCR Antagonistic Peptides

As used herein, the terms “peptide” and “polypeptide” include linear,branched and cyclic peptides. As used herein, a CXC receptor (CXCR)refers to any chemokine receptor that specifically binds and responds toa cytokine of the CXC chemokine family. A CXCR includes, but is notlimited to, CXCR1, CXXR2, CXCR2, CXCR3, CXCR4, CXCR5, CXCR5, CXCR6,and/or CXCR7. As used herein, the term “CXCR antagonistic peptide”refers to any peptide that has a negative impact in a function of aCXCR. In some embodiments, a CXCR antagonistic peptide directly orindirectly reduces, inhibits, or down-regulates the activity of a CXCR.In some embodiments, provided peptides are isolated peptides.

According to the present invention, a CXCR antagonistic peptide containsthe amino acid sequence proline-glycine-X (PGX, or Pro-Gly-Xaa), whereinX is any amino acid. In some embodiments, X is selected from Alanine,arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid,glycine, histidine, isoleucine, leucine, lysine, methionine,phenylalanine, proline, serine, threonine, tryptophan, tyrosine orvaline. In some embodiments, X is not proline. In some embodiments, X isglycine.

According to various embodiments, amino acids may be from natural ornon-natural sources, provided that at least one amino group and at leastone carboxyl group are present in the molecule. In some embodiments,provided peptides comprise naturally occurring amino acids. In someembodiments, provided peptides comprise one or more synthetic orun-naturally occurring amino acids (e.g., α,α-disubstituted amino acids,N-alkyl amino acids). In some embodiments, provided peptides compriseone or more D-amino acid. In some embodiments, provided peptidescomprise one or more L-amino acid. Provided peptides may also containone or more rare amino acids (such as 4-hydroxyproline orhydroxylysine), organic acids or amides and/or derivatives of commonamino acids, such as amino acids having the C-terminal carboxylateesterified (e.g., benzyl, methyl or ethyl ester) or amidated and/orhaving modifications, for example, of the N-terminal amino group (e.g.,acetylation or alkoxycarbonylation), with or without any of a widevariety of side-chain modifications and/or substitutions (e.g.,methylation, benzylation, t-butylation, tosylation, alkoxycarbonylation,and the like). Exemplary modifications are further described below.

In some embodiments, provided peptides comprise one or more residuesother than common amino acids. According to various embodiments,residues other than common amino acids that may be present in a providedpeptide include, but are not limited to, penicillamine,β,β-tetramethylene cysteine, β,β-pentamethylene cysteine,β-mercaptopropionic acid, β,β-pentamethylene-β-mercaptopropionic acid,2-mercaptobenzene, 2-mercaptoaniline, 2-mercaptoproline, ornithine,diaminobutyric acid, α-aminoadipic acid, m-aminomethylbenzoic acid andα,β-diaminopropionic acid.

In some embodiments, the present invention provides polypeptidescomprising an amino acid sequence of between 3 and 60 amino acids inlength (e.g., between 3-55, 3-50, 3-45, 3-40, 3-35, 3-30, 3-25, 3-20,3-15, 3-10, or 3-6 amino acids in length). In some embodiments, providedpeptides are between 3 and 15 amino acids in length (e.g., 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in length). In someembodiments, provided peptides are less than 15 amino acids in length(e.g., less than 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, or 4 amino acids inlength). In some embodiments, provided peptides are less than 6 aminoacids in length.

In some embodiments, the sequence Pro-Gly-Xaa is located at theN-terminus of the peptide. In some embodiments, the sequence Pro-Gly-Xaais located at the C-terminus of the peptide. In some embodiments, thesequence Pro-Gly-Xaa is located internally.

In some embodiments, the sequence Pro-Gly-Xaa may appear as repeats. Forexample, a peptide according to the present invention may contain 2-50,2-40, 2-30, 2-20, 2-15, 2-10, 2-8, 2-6, or 2-5 PGX repeats. In someembodiments, a peptide according to the present invention may containmore than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30,35, 40, 45 or 50 PGX repeats. The PGX repeats may appear as tandemrepeats (i.e., consecutive repeats), may be interrupted by non-PGXsequences (e.g., by one or more amino acids that do not appear as PGX),or both. the PGX repeats can also be linear, branched or both. In someembodiments, Xaa comprises the same amino acid each repeat. In someembodiments, Xaa comprises a different amino acid (and/or a differentform of a particular amino acid) in each repeat. In some embodiments,Xaa comprises a different amino acid (and/or a different form of aparticular amino acid) in two or more repeats.

Peptide Modifications

In some embodiments, amino acids at one or more positions in a providedpeptide comprise one or more modifications. In some embodiments, aparticular modification may impart one or more improved characteristicsto a peptide. For example, a modification may increase the potencyand/or half-life of a peptide.

In some embodiments, provided peptides are modified at the N-terminus.For example, N-terminal acetylation, methylation, oralkoxycarbonylation. In some embodiments, a peptide according to theinvention is N-acetylated PGX, wherein X is any amino acid. In someembodiments, X is not proline. In particular embodiments, a peptideaccording to the invention is N-acetylated PGG. In some embodiments,provided proteins are modified at the C-terminus. In some embodiments,provided peptides are modified at an amino acid position other than theN-terminal or C-terminal position.

Any of a variety of known modifications of amino acids may beimplemented in the scope of the invention. For example, several knownmodifications compatible with various embodiments may be found anddescribed atwww.piercenet.com/previews/guides/custom-peptide-modifications, thedisclosure of which is hereby incorporate by reference. In someembodiments, a modification comprises attachment of an acetyl group to aparticular amino acid. In some embodiments, a modification is PEGylationcomprising the addition of at least one monomeric unit of polyethyleneglycol (PEG):

In some embodiments, provided peptides comprise two or more monomericunits of PEG (i.e. PEG oligomers or multimers).

In some embodiments, a modification comprises hydroxylation, oxidation,phosphorylation, adenylylation, glycation, glycosylation, and/orbiotinylation of a particular amino acid.

In some embodiments, provided peptides are modified by the attachment ofa alkyl group (e.g. methyl, ethyl) to one or more amino acids. In someembodiments, provided peptides are modified by the attachment of a alkylgroup to one or more amino acids. In some embodiments, provided peptidesare modified by the attachment of a glycosyl group to one or more aminoacids. In some embodiments, provided peptides are modified by theattachment of a flavin and/or 4′-phosphopantetheinyl moiety to one ormore amino acids.

In some embodiments, provided peptides are modified by citrullination,deamidation, ubiquitination, eliminylation, and/or carbamylation. Insome embodiments, provided peptides are modified by the attachment ofone or more myristate, lipoate, palmitate, and/or isoprenoid molecules.

In some embodiments, provided peptides are modified by pyroglutamateformation. In some embodiments, provided peptides are modified byS-glutathionylation. In some embodiments, provided peptides are modifiedby S-nitrosylation. In some embodiments, provided peptides are modifiedby succinylation. In some embodiments, provided peptides are modified bysulfation.

Cyclic Peptides

In some embodiments, provided peptide are cyclic peptides. As usedherein, a cyclic peptide has an intramolecular covalent bond between twonon-adjacent residues. The intramolecular bond may be a backbone tobackbone, side-chain to backbone or side-chain to side-chain bond (i.e.,terminal functional groups of a linear peptide and/or side-chainfunctional groups of a terminal or interior residue may be linked toachieve cyclization). Typical intramolecular bonds include disulfide,amide and thioether bonds. A variety of means for cyclizing polypeptidesare well known in the art, as are many other modifications that can bemade to such peptides. For a general discussion, see InternationalPatent Publication Nos. WO 01/53331 and WO 98/02452, the contents ofwhich are incorporated herein by reference. Such cyclic bonds and othermodifications can also be applied to the cyclic peptides and derivativecompounds of this invention.

As with provided linear or branched peptides, amino acids in providedcyclic peptides may comprise one or more modifications. In someembodiments, amino acids may be from natural or non-natural sources,provided that at least one amino group and at least one carboxyl groupare present in the molecule; α- and β-amino acids are generallypreferred. Cyclic peptides may also contain one or more rare amino acids(such as 4-hydroxyproline or hydroxylysine), organic acids or amidesand/or derivatives of common amino acids, such as amino acids having theC-terminal carboxylate esterified (e.g., benzyl, methyl or ethyl ester)or amidated and/or having modifications of the N-terminal amino group(e.g., acetylation or alkoxycarbonylation), with or without any of awide variety of side-chain modifications and/or substitutions (e.g.,methylation, benzylation, t-butylation, tosylation, alkoxycarbonylation,and the like). Suitable derivatives include amino acids having anN-acetyl group (such that the amino group that represents the N-terminusof the linear peptide prior to cyclization is acetylated) and/or aC-terminal amide group (i.e., the carboxy terminus of the linear peptideprior to cyclization is amidated). Residues other than common aminoacids that may be present with a cyclic peptide include, but are notlimited to, penicillamine, β,β-tetramethylene cysteine,β,β-pentamethylene cysteine, β-mercaptopropionic acid,β,β-pentamethylene-β-mercaptopropionic acid, 2-mercaptobenzene,2-mercaptoaniline, 2-mercaptoproline, ornithine, diaminobutyric acid,α-aminoadipic acid, m-aminomethylbenzoic acid and α,β-diaminopropionicacid.

Following synthesis of a linear peptide, with or without N-acetylationand/or C-amidation, cyclization may be achieved by any of a variety oftechniques well known in the art. Within one embodiment, a bond may begenerated between reactive amino acid side chains. For example, adisulfide bridge may be formed from a linear peptide comprising twothiol-containing residues by oxidizing the peptide using any of avariety of methods. Within one such method, air oxidation of thiols cangenerate disulfide linkages over a period of several days using eitherbasic or neutral aqueous media. The peptide is used in high dilution tominimize aggregation and intermolecular side reactions. Alternatively,strong oxidizing agents such as I2 and K3Fe(CN)6 can be used to formdisulfide linkages. Those of ordinary skill in the art will recognizethat care must be taken not to oxidize the sensitive side chains of Met,Tyr, Trp or His. Within further embodiments, cyclization may be achievedby amide bond formation. For example, a peptide bond may be formedbetween terminal functional groups (i.e., the amino and carboxy terminiof a linear peptide prior to cyclization). Alternatively, cyclizationmay be accomplished by linking one terminus and a residue side chain orusing two side chains, with or without an N-terminal acetyl group and/ora C-terminal amide. Exemplary residues capable of forming a lactam bondinclude lysine, ornithine (Orn), α-amino adipic acid,m-aminomethylbenzoic acid, α,β-diaminopropionic acid, glutamate oraspartate. Methods for forming amide bonds are generally well known inthe art. Within one such method, carbodiimide-mediated lactam formationcan be accomplished by reaction of the carboxylic acid with DCC, DIC, EDAC or DCCI, resulting in the formation of an O-acylurea that can bereacted immediately with the free amino group to complete thecyclization. Alternatively, cyclization can be performed using the azidemethod, in which a reactive azide intermediate is generated from analkyl ester via a hydrazide. Alternatively, cyclization can beaccomplished using activated esters. The presence of electronwithdrawing substituents on the alkoxy carbon of esters increases theirsusceptibility to aminolysis. The high reactivity of esters ofp-nitrophenol, N-hydroxy compounds and polyhalogenated phenols has madethese “active esters” useful in the synthesis of amide bonds. Within afurther embodiment, a thioether linkage may be formed between the sidechain of a thiol-containing residue and an appropriately derivatizedα-amino acid. By way of example, a lysine side chain can be coupled tobromoacetic acid through the carbodiimide coupling method (DCC, EDAC)and then reacted with the side chain of any of the thiol containingresidues mentioned above to form a thioether linkage. In order to formdithioethers, any two thiol containing side-chains can be reacted withdibromoethane and diisopropylamine in DMF.

Therapeutic Application

Various peptides described herein may be used to inhibit, reduce,decrease, or down-regulate a CXCR activity in a cell or a subject. Forexample, a peptide described herein may be used to inhibit, reduce,decrease, or down-regulate a CXCR activity and/or a CXCR mediatedpathway in a mammalian cell (e.g., a human cell), such as in a culturedcell or a cell of a living organism. In some embodiments, peptidesdescribed herein can also be used to inhibit, reduce, decrease, ordown-regulate a CXCR activity in a subject. In some embodiments, apeptide described herein inhibits, reduces, decreases, or down-regulatesthe activity of CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, and/or CXCR7.In particular embodiments, a peptide described herein inhibits, reduces,decreases, or down-regulates the activity of CXCR1 and/or CXCR2.

In various embodiments, peptides described herein may be used to treatdiseases, disorders and conditions in which a CXCR mediated pathway isimplicated. In some embodiments, peptides described herein may be usedto treat inflammation, stroke, traumatic brain injury, pancreaticcancer, to name but a few.

Exemplary diseases, disorders, or conditions that can be treatedaccording to the present invention are provided below.

Inflammation

In some embodiments, provided peptides are used in the treatment ofinflammation. For example, provided peptides can be used in thetreatment of inflammation associated with arthritis (includingrheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis,psoriatic arthritis, lupus-associated arthritis or ankylosingspondylitis); scleroderma; systemic lupus erythematosis; HIV; Sjogren'ssyndrome; vasculitis; multiple sclerosis; autoimmune thyroiditis; asthma(e.g., allergic and non-allergic asthma); dermatitis (including atopicdermatitis and eczematous dermatitis); myasthenia gravis; inflammatorybowel disease (IBD); Crohn's disease; colitis; diabetes mellitus (typeI); inflammatory conditions of, e.g., the skin (e.g., psoriasis),cardiovascular system (e.g., atherosclerosis), nervous system (e.g.,Alzheimer's disease), liver (e.g., hepatitis), kidney (e.g., nephritis)and pancreas (e.g., pancreatitis); sarcoidosis; scleroderma; cirrhosis;eosinophilic esophagitis; cardiovascular disorders (e.g., cholesterolmetabolic disorders, oxygen free radical injury, ischemia, pulmonaryfibrosis, idiopathic pulmonary fibrosis, reperfusion injury, acutevaso-occlusive crisis in sickle cell anemia; disorders associated withwound healing; respiratory disorders, e.g., asthma, COPD (e.g., cysticfibrosis); acute inflammatory conditions (e.g., endotoxemia, sepsis andsepticaemia, toxic shock syndrome and infectious disease (e.g.,myocarditis, cardiomyopathy, acute endocarditis, pericarditis); SystemicInflammatory Response Syndrome (SIRS)/sepsis; atopic disorders, e.g.,urticaria, allergic rhinitis, rhinosinusitis (e.g., chronic allergicrhinosinusitis) allergic enterogastritis; adult respiratory distresssyndrome (ARDS); systemic erythematosis (SLE); Airwayhyperresponsiveness (AHR); bronchial hyperreactivity; ChronicObstructive Pulmonary disease (COPD); Congestive Heart Failure (CHF);inflammatory bowel disease; inflammatory complications of diabetesmellitus; metabolic syndrome; end-stage renal disease (ESRD); musclefatigue or inflammation and dermal conditions; inflammatory conditionscaused by bacterial infection or viral infection (e.g. respiratorysyncytial virus-induced airway hyperreactivity); tumors or cancers(e.g., soft tissue or solid tumors), such as leukemia (e.g., Hodgkin'slymphoma), glioblastoma, astrocytoma, lymphoma, melanoma or prostatecancer; or transplant rejection (e.g. destruction of pancreatic isletcells in islet cell transplantation, delayed graft failure in kidney orother organ transplantation); acute and chronic graft versus hostdisease.

Arthritis

In some embodiments, provided peptides are used in the treatment ofarthritis (including rheumatoid arthritis, juvenile rheumatoidarthritis, osteoarthritis, psoriatic arthritis, lupus-associatedarthritis or ankylosing spondylitis). Arthritis is a joint disorder thatinvolves inflammation of one or more joints. There are over 100different forms of arthritis. The most common form, osteoarthritis(degenerative joint disease), is a result of trauma to the joint,infection of the joint, or age. Other arthritis forms are rheumatoidarthritis, psoriatic arthritis, and related autoimmune diseases. Septicarthritis is caused by joint infection.

The major complaint by individuals who have arthritis is joint pain.Pain is often a constant and may be localized to the joint affected. Thepain from arthritis is due to inflammation that occurs around the joint,damage to the joint from disease, daily wear and tear of joint, musclestrains caused by forceful movements against stiff painful joints andfatigue. Other common symptoms of arthritis include varied levels ofswelling, joint stiffness and a constant ache around the joint(s).

Arthritis is the most common cause of disability in the USA. More than20 million individuals with arthritis have severe limitations infunction on a daily basis. Absenteeism and frequent visits to thephysician are common in individuals who have arthritis. Arthritis makesit very difficult for individuals to be physically active and somebecome home bound. It is estimated that the total cost of arthritiscases is close to $100 billion of which nearly 50% is from lostearnings. Each year, arthritis results in nearly 1 millionhospitalizations and close to 45 million outpatient visits to healthcare centers. Arthritis can make it very difficult for an individual toremain physically active, contributing to an increased risk of obesity,high cholesterol or vulnerability to heart disease. Individuals witharthritis are also at increased risk of depression, which may be relatedto fear of worsening symptoms. Other significant secondary changes thatare common in patients with advanced arthritis are muscle weakness, lossof flexibility and decreased aerobic fitness, which can all occur inpatients who have limited their physical activity.

Rheumatoid Arthritis

In some embodiments, provided peptides are used in the treatment ofrheumatoid arthritis. Rheumatoid arthritis is a chronic inflammatorydisorder that typically affects the small joints in the hands and feet.Unlike the wear-and-tear damage of osteoarthritis, rheumatoid arthritisaffects the lining of the joints, causing a painful swelling that caneventually result in bone erosion and joint deformity.

Signs and symptoms of rheumatoid arthritis may include: tender, warm,swollen joints, morning stiffness that may last for hours, firm bumps oftissue under the skin on the arms (rheumatoid nodules), fatigue, feverand weight loss. Early rheumatoid arthritis tends to affect the smallerjoints first, particularly the joints that attach the fingers to thehands and the toes to the feet. As the disease progresses, symptomsoften spread to the knees, ankles, elbows, hips and shoulders. In mostcases, symptoms occur in the same joints on both sides of the body.Rheumatoid arthritis signs and symptoms may vary in severity and mayeven come and go. Periods of increased disease activity, called flares,alternate with periods of relative remission, when the swelling and painfade or disappear. Over time, rheumatoid arthritis can cause joints todeform and shift out of place.

Rheumatoid arthritis occurs when the immune system attacks the synovium,the lining of the membranes that surround joints. The resultinginflammation thickens the synovium, which can eventually destroy thecartilage and bone within the joint. The tendons and ligaments that holdthe joint together weaken and stretch. Gradually, the joint loses itsshape and alignment. Doctors don't know what starts this process,although a genetic component appears likely. While one's genes don'tactually cause rheumatoid arthritis, they can make a person moresusceptible to environmental factors, such as infection with certainviruses and bacteria, which may trigger the disease.

Juvenile Rheumatoid Arthritis

In some embodiments, provided peptides are used in the treatment ofjuvenile rheumatoid arthritis. Juvenile rheumatoid arthritis is the mostcommon type of arthritis in children under the age of 16. Juvenilerheumatoid arthritis causes persistent joint pain, swelling andstiffness. In some cases, juvenile arthritis affects the entire body,causing swollen lymph nodes, rashes and fever. Some children mayexperience symptoms for only a few months, while others have symptomsfor the rest of their lives.

Some types of juvenile rheumatoid arthritis can cause seriouscomplications, such as growth problems and eye inflammation (uveitis).If eye inflammation is left untreated, it may result in cataracts,glaucoma and even blindness. Eye inflammation frequently occurs withoutsymptoms, so it's important for children with juvenile rheumatoidarthritis to be examined regularly by an ophthalmologist. Growthproblems associated with juvenile rheumatoid arthritis can also involveproblems with bone development; some medications used to treat thedisease, such as corticosteroids, can also inhibit growth.

Juvenile rheumatoid arthritis occurs when the body's immune systemattacks its own cells and tissues. It's unknown why this happens, butboth heredity and environment seem to play a role. Certain genemutations may make a person more susceptible to environmentalfactors—such as viruses—that may trigger the disease.

Osteoarthritis

In some embodiments, provided peptides are used in the treatment ofosteoarthritis. Osteoarthritis is one of the most common form ofarthritis. It can affect both the larger and the smaller joints of thebody, including the hands, wrists, feet, back, hip, and knee. Thedisease is essentially one acquired from daily wear and tear of thejoint; however, osteoarthritis can also occur as a result of injury.Osteoarthritis begins in the cartilage and eventually causes the twoopposing bones to erode into each other.

Initially, the condition starts with minor pain during activities, butsoon the pain can be continuous and even occur while in a state of rest.The pain can be debilitating and prevent one from doing some activities.Other symptoms can include tenderness (joints may feel tender when lightpressure is applied), stiffness (most noticeable after awakening orafter a period of inactivity), loss of flexibility, a grating sensationand bone spurs (extra bits of bone that feel like hard lumps and canform around the affected joint). Osteoarthritis typically affects theweight-bearing joints, such as the back, spine, and pelvis.

Unlike rheumatoid arthritis, osteoarthritis is most commonly a diseaseof the elderly. More than 30 percent of women have some degree ofosteoarthritis by age 65. Osteoarthritis occurs when the cartilage thatcushions the ends of bones in joints deteriorates over time. Cartilageis a firm, slippery tissue that permits nearly frictionless jointmotion. In osteoarthritis, the slick surface of the cartilage becomesrough. Eventually, if the cartilage wears down completely, a patient maybe left with bone rubbing on bone. Risk factors for osteoarthritisinclude prior joint trauma, obesity, and a sedentary lifestyle.

Psoriatic Arthritis

In some embodiments, provided peptides are used in the treatment ofpsoriatic arthritis. Psoriatic arthritis is a form of arthritis thataffects some people who have psoriasis, a condition that features redpatches of skin topped with silvery scales. Most people developpsoriasis first and are later diagnosed with psoriatic arthritis, butthe joint problems can sometimes begin before skin lesions appear. Jointpain, stiffness and swelling are the main symptoms of psoriaticarthritis.

They can affect any part of the body, including fingertips and spine,and can range from relatively mild to severe. Psoriatic arthritis cancause a painful, sausage-like swelling of the fingers and toes. Patientsmay also develop swelling and deformities in the hands and feet beforehaving significant joint symptoms. Psoriatic arthritis can also causepain at the points where tendons and ligaments attach to thebones—especially at the back of the heel (Achilles tendinitis) or in thesole of the foot (plantar fasciitis). Some people develop a conditioncalled spondylitis as a result of psoriatic arthritis. Spondylitismainly causes inflammation of the joints between the vertebrae of thespine and in the joints between the spine and pelvis (sacroiliitis). Asmall percentage of people with psoriatic arthritis develop arthritismutilans—a severe, painful and disabling form of the disease. Over time,arthritis mutilans destroys the small bones in the hands, especially thefingers, leading to permanent deformity and disability. In bothpsoriasis and psoriatic arthritis, disease flares may alternate withperiods of remission.

Psoriatic arthritis occurs when the body's immune system begins toattack healthy cells and tissue. The abnormal immune response causesinflammation in the joints as well as overproduction of skin cells. It'snot entirely clear why the immune system turns on healthy tissue, but itseems likely that both genetic and environmental factors play a role.Many people with psoriatic arthritis have a family history of eitherpsoriasis or psoriatic arthritis. Researchers have discovered certaingenetic markers that appear to be associated with psoriatic arthritis.Physical trauma or something in the environment, such as a viral orbacterial infection, may trigger psoriatic arthritis in people with aninherited tendency.

Ankylosing Spondylitis

In some embodiments, provided peptides are used in the treatment ofankylosing spondylitis. Ankylosing spondylitis is an inflammatorydisease that can cause some of the vertebrae in the spine to fusetogether. This fusing makes the spine less flexible and can result in ahunched-forward posture. A severe case of ankylosing spondylitis canmake it impossible for someone to lift their head high enough to seeforward. Ankylosing spondylitis affects men more often than women.

Early signs and symptoms of ankylosing spondylitis may include pain andstiffness in the lower back and hips, especially in the morning andafter periods of inactivity. These symptoms may come on so graduallythat one doesn't notice them at first. Over time, symptoms may worsen,improve or stop completely at irregular intervals. The areas of the bodymost commonly affected are: the joint between the base of the spine andthe pelvis, the vertebrae in the lower back, the places where tendonsand ligaments attach to bones, mainly in the spine, but sometimes alongthe back of the heel, the cartilage between the breastbone and ribs, hipand shoulder joints.

Ankylosing spondylitis has no known specific cause, though geneticfactors seem to be involved. In particular, people who have a genecalled HLA-B27 are at significantly increased risk of developingankylosing spondylitis. As ankylosing spondylitis worsens and theinflammation persists, new bone forms as part of the body's attempt toheal. This new bone gradually bridges the gap between vertebrae andeventually fuses sections of vertebrae together. Those parts of thespine become stiff and inflexible. Fusion can also stiffen the rib cage,restricting lung capacity and function.

Vasulitis

In some embodiments, provided peptides are used in the treatment ofvasculitis. Vasculitis is an inflammation of the blood vessels.Vasculitis causes changes in the walls of the blood vessels, includingthickening, weakening, narrowing and scarring. There are many types ofvasculitis. Some forms last only a short time (acute) while others arelong lasting (chronic). Vasculitis, which is also known as angiitis andarteritis, can be so severe that the tissues and organs supplied by theaffected vessels don't get enough blood. This shortage of blood canresult in organ and tissue damage, even death. Vasculitis can affectanyone, though some types of vasculitis are more common among certaingroups. Some forms of vasculitis improve on their own, but othersrequire treatment—often including taking medications for an extendedperiod of time.

The signs and symptoms of vasculitis vary depending on which bloodvessels and, as a result, which organ systems are affected. However,general signs and symptoms that many people with vasculitis experienceinclude: fever, fatigue, weight loss, muscle and joint pain, loss ofappetite, and nerve problems, such as numbness or weakness. Signs andsymptoms for some types of vasculitis include:

Behcet's syndrome: This condition causes inflammation of the arteriesand veins, and often appears in people in their 20s and 30s. Signs andsymptoms include mouth and genital ulcers, eye inflammation, andacne-like lesions on the skin.

Buerger's disease: Also called thromboangiitis obliterans, thiscondition causes inflammation and clots in the blood vessels in theextremities. Signs and symptoms can include pain in the hands, arms,feet and legs, and ulcers on the fingers and toes. This disorder isstrongly associated with cigarette smoking.

Churg-Strauss syndrome: This condition, also known as allergicgranulomatosis and allergic angiitis, most commonly affects the bloodvessels in the lungs. It's often associated with asthma.

Cryoglobulinemia: This condition is often associated with hepatitis Cinfections. Signs and symptoms include a rash called purpura on thelower extremities, arthritis, weakness and nerve damage (neuropathy).

Giant cell arteritis: This condition, which usually occurs in peopleolder than 50, is an inflammation of the arteries in the head,especially the temples. Giant cell arteritis can cause headaches, scalptenderness, jaw pain while chewing, blurred or double vision, and evenblindness. Giant cell arteritis is often associated with another type ofinflammatory condition called polymyalgia rheumatica (PMR). PMR causespain in and inflammation of the large joints, such as the shoulders andhips. Signs and symptoms include pain and stiffness in the muscles ofthe hips, thighs, shoulders, upper arms and neck.

Henoch-Schonlein purpura: This condition is caused by inflammation ofthe blood vessels of the skin, joints, bowel and kidneys. Signs andsymptoms can include abdominal pain, blood in the urine, joint pain, anda rash called purpura on the buttocks, legs and feet. Henoch-Schonleinusually affects children, but it can occur at any age.

Hypersensitivity vasculitis: The primary sign of hypersensitivityvasculitis is red spots on the skin. It can be triggered by an allergy,most often to a medication or an infection.

Kawasaki disease: Also known as mucocutaneous lymph node syndrome, thiscondition most often affects children younger than 5 years of age. Signsand symptoms include fever, skin rash and eye inflammation.

Microscopic polyangiitis: This form of vasculitis affects small-sizedblood vessels in the kidneys, lungs and skin. Signs and symptoms includeskin lesions, fever, unintentional weight loss,glomerulonephritis—inflammation of the small blood vessels in thekidneys—and nerve damage.

Polyarteritis nodosa: This form of vasculitis affects medium-sized bloodvessels in many different parts of the body, including the skin, heart,kidneys, peripheral nerves, muscles and intestines. Signs and symptomsinclude a rash called purpura, skin ulcers, muscle and joint pain,abdominal pain, and kidney problems.

Takayasu's arteritis: This form of vasculitis includes the largestarteries in the body, including the aorta, and typically occurs in youngwomen. Signs and symptoms include a feeling of numbness or cold in theextremities, decreased or absent pulses, high blood pressure, headaches,and visual disturbances.

Granulomatosis with polyangiitis (Wegener's): Formerly known asWegener's granulomatosis, granulomatosis with polyangiitis causesinflammation of the blood vessels in the nose, sinuses, throat, lungsand kidneys. Signs and symptoms can include nasal stuffiness, chronicsinus infections and nosebleeds. The kidneys are often affected, thoughmost people won't have any noticeable symptoms until the damage is moreadvanced.

Vasculitis occurs when the immune system mistakenly sees blood vesselcells as foreign. The immune system then attacks those cells as if theywere an invader, such as a bacteria or virus. It's not always clear whythis happens, but an infection, some cancers, certain immune systemdisorders or an allergic reaction may serve as the trigger. Bloodvessels affected by vasculitis become inflamed, which can cause thelayers of the blood vessel wall to thicken. This narrows the bloodvessels, reducing the amount of blood—and therefore oxygen and vitalnutrients—that reaches the body's tissues. In some cases, a blood clotmay form in an affected blood vessel, obstructing blood flow. Sometimesinstead of becoming narrower, a blood vessel may weaken and form a bulge(aneurysm), a potentially life-threatening condition. For many of itsforms, the cause of vasculitis is unknown. These forms of vasculitis arecalled primary vasculitis. Forms of vasculitis for which an underlyingdisease is the cause are called secondary vasculitis. Examples of causesof secondary vasculitis include: infections, immune system diseases,allergic reactions, and blood cell cancers.

Complications of vasculitis depend on the type of vasculitis a personhas. In general, complications that can occur include: organ damage(some types of vasculitis can be severe, causing damage to major organsand recurring episodes of vasculitis (Even when treatment for vasculitisis initially successful, the condition may recur and require additionaltreatment; in other cases, vasculitis may never completely go away andrequires ongoing treatment).

Multiple Sclerosis

In some embodiments, provided peptides are used in the treatment ofmultiple sclerosis (MS). Multiple sclerosis is a potentiallydebilitating disease in which the body's immune system eats away at theprotective sheath (myelin) that covers the nerves. Damage to myelincauses interference in the communication between the brain, spinal cordand other areas of the body. This condition may result in deteriorationof the nerves themselves, a process that's not reversible.

Symptoms of multiple sclerosis vary, depending on the location ofaffected nerve fibers. Multiple sclerosis symptoms may include: numbnessor weakness in one or more limbs, partial or complete loss of centralvision, usually in one eye, often with pain during eye movement (opticneuritis), double vision or blurring of vision, tingling or pain inparts of the body, electric-shock sensations that occur with certainhead movements, tremor, lack of coordination or unsteady gait, slurredspeech, fatigue, and dizziness. Heat sensitivity is common in peoplewith multiple sclerosis. Small increases in body temperature can triggeror worsen multiple sclerosis symptoms. Most people with multiplesclerosis, particularly in the beginning stages of the disease,experience relapses of symptoms, which are followed by periods ofcomplete or partial remission of symptoms. Some people have a benignform of multiple sclerosis. In this form of the disease, the conditionremains stable and often doesn't progress to serious forms of MS afterthe initial attack. In some cases, people with multiple sclerosis mayalso develop: muscle stiffness of spasms, paralysis, most typically inthe legs, problems with bladder, bowel or sexual function, mentalchanges, such as forgetfulness or difficulties concentrating, depressionand epilepsy.

The cause of multiple sclerosis is unknown. It's believed to be anautoimmune disease, in which the body's immune system attacks its owntissues. In multiple sclerosis, this process destroys myelin—the fattysubstance that coats and protects nerve fibers in the brain and spinalcord. Myelin can be compared to the insulation on electrical wires. Whenmyelin is damaged, the messages that travel along that nerve may beslowed or blocked. Doctors and researchers don't understand why multiplesclerosis develops in some people and not others. A combination offactors, ranging from genetics to childhood infections, may play a role.

Inflammatory Bowel Disease (IBD)

In some embodiments, provided peptides are used in the treatment ofinflammatory bowel disease (IBD). Inflammatory bowel disease (IBD)involves chronic inflammation of all or part of the digestive tract. IBDprimarily includes ulcerative colitis and Crohn's disease. IBD can bepainful and debilitating, and sometimes leads to life-threateningcomplications. Ulcerative colitis is an inflammatory bowel disease thatcauses long-lasting inflammation in part of the digestive tract.Symptoms usually develop over time, rather than suddenly. Ulcerativecolitis usually affects only the innermost lining of the large intestine(colon) and rectum. It occurs only through continuous stretches of thecolon. Crohn's disease is an inflammatory bowel disease that causesinflammation anywhere along the lining of the digestive tract, and oftenspreads deep into affected tissues. This can lead to abdominal pain,severe diarrhea and even malnutrition. The inflammation caused byCrohn's disease can involve different areas of the digestive tract indifferent people. Collagenous colitis and lymphocytic colitis also areconsidered inflammatory bowel diseases, but are usually regardedseparately from classic inflammatory bowel disease.

Inflammatory bowel disease symptoms vary, depending on the severity ofinflammation and where it occurs. Ulcerative colitis is classifiedaccording to its signs and symptoms. In ulcerative proctitis,inflammation is confined to the area closest to the anus (rectum), andfor some people, rectal bleeding may be the only sign of the disease.Others may have rectal pain, a feeling of urgency or have frequent,small bowel movements. This form of ulcerative colitis tends to be themildest. In proctosigmoiditis, the rectum and the lower end of thecolon, known as the sigmoid colon are involved. Bloody diarrhea,abdominal cramps and pain, and an inability to move the bowels in spiteof the urge to do so (tenesmus) are common problems associated with thisform of the disease. In left-sided colitis, inflammation extends fromthe rectum up through the sigmoid and descending colon, which arelocated in the upper left part of the abdomen. Signs and symptomsinclude bloody diarrhea, abdominal cramping and pain on the left side,and unintended weight loss. In pancolitis, more than the left colon, andoften the entire colon, are affected. Pancolitis causes bouts of bloodydiarrhea that may be severe, abdominal cramps and pain, fatigue, andsignificant weight loss. Fulminant colitis is a rare, life-threateningform of colitis that affects the entire colon and causes severe pain,profuse diarrhea and, sometimes, dehydration and shock. People withfulminant colitis are at risk of serious complications, including colonrupture and toxic megacolon, a condition that causes the colon torapidly expand. The course of ulcerative colitis varies, with periods ofacute illness often alternating with periods of remission. Most peoplewith a milder condition, such as ulcerative proctitis, won't go on todevelop more severe signs and symptoms. IBD may lead to one or more ofthe following complications: bowel obstruction, ulcers, fistulas, analfissure, malnutrition, colon cancer, and other health problems such asarthritis, inflammation of eyes or skin, clubbing of fingernails, kidneystones, gallstones, inflammation of the bile ducts, and osteoporosis.

Crohn's Disease

In some embodiments, provided peptides are used in the treatment ofCrohn's disease. Crohn's disease is an inflammatory bowel disease (IBD).It causes inflammation of the lining of the digestive tract, which canlead to abdominal pain, severe diarrhea and even malnutrition.Inflammation caused by Crohn's disease can involve different areas ofthe digestive tract in different people. The inflammation caused byCrohn's disease often spreads deep into the layers of affected boweltissue. Like ulcerative colitis, another common IBD, Crohn's disease canbe both painful and debilitating, and sometimes may lead tolife-threatening complications.

Inflammation of Crohn's disease may involve different areas in differentpeople. In some people, just the small intestine is affected. In others,it's confined to the colon (part of the large intestine). The mostcommon areas affected by Crohn's disease are the last part of the smallintestine (ileum) and the colon. Inflammation may be confined to thebowel wall, which can lead to scarring (stenosis), or inflammation mayspread through the bowel wall (fistula). Signs and symptoms of Crohn'sdisease can range from mild to severe and may develop gradually or comeon suddenly, without warning. A person may also have periods of timewhen they have no signs or symptoms (remission). When the disease isactive, signs and symptoms may include: diarrhea, abdominal pain andcramping (may include nausea and vomiting), bloody stool, ulcers, andreduced appetite and weight loss. Others signs and symptoms include:fever, arthritis, eye inflammation, mouth sores, skin disorders,inflammation of the liver or bile ducts, and, in children, delayedgrowth or sexual development. Additionally, Crohn's disease may lead toone or more of the following complications: bowel obstruction, ulcers,fistulas, anal fissure, malnutrition, color cancer, and other healthproblems such as arthritis, inflammation of eyes or skin, clubbing offingernails, kidney stones, gallstones, inflammation of the bile ducts,and osteoporosis.

Organ System Specific Inflammation

In some embodiments, provided peptides are used in the treatment ofinflammation associated with inflammatory conditions of the skin (e.g.,psoriasis), cardiovascular system (e.g., atherosclerosis), nervoussystem (e.g., Alzheimer's disease), liver (e.g., hepatitis), kidney(e.g., nephritis) and pancreas (e.g., pancreatitis).

Nervous System

In some embodiments, provided peptides are used in the treatment ofinflammation associated with inflammatory conditions of the nervoussystem (e.g. Alzheimer's disease). Alzheimer's disease is a progressivedisease that destroys memory and other important mental functions. Ifsthe most common cause of dementia—a group of brain disorders thatresults in the loss of intellectual and social skills. These changes aresevere enough to interfere with day-to-day life. In Alzheimer's disease,the connections between brain cells and the brain cells themselvesdegenerate and die, causing a steady decline in memory and mentalfunction.

At first, increasing forgetfulness or mild confusion may be the onlysymptoms of Alzheimer's disease that are noticed. But over time, thedisease robs a person of more of their memory, especially recentmemories. The rate at which symptoms worsen varies from person toperson. In a person with Alzheimer's, unusual difficulty rememberingthings and organizing thoughts may be the first things they notice. Onthe other hand, some people with Alzheimer's may not recognize thatanything is wrong, even when changes are noticeable to family members,close friends or co-workers. Brain changes associated with Alzheimer'sdisease lead to growing trouble with memory, disorientation andmisinterpreting spatial relationships, speaking and writing, thinkingand reasoning, making judgments and decisions, planning and performingfamiliar tasks and changes in personality and behavior. Memory loss,impaired judgment and other cognitive changes caused by Alzheimer's cancomplicate treatment for other health conditions. A person withAlzheimer's disease may not be able to: communicate that he or she isexperiencing pain—for example, from a dental problem, report symptoms ofanother illness, follow a prescribed treatment plan, or notice ordescribe medication side effects. As Alzheimer's disease progresses tolater stages, brain changes begin to affect physical functions, such asswallowing, balance, and bowel and bladder control. These effects canincrease vulnerability to additional health problems such as: pneumoniaand other infections and injuries from falls.

Scientists believe that for most people, Alzheimer's disease resultsfrom a combination of genetic, lifestyle and environmental factors thataffect the brain over time. Less than 5 percent of the time, Alzheimer'sis caused by specific genetic changes that virtually guarantee a personwill develop the disease. Although the causes of Alzheimer's are not yetfully understood, its effect on the brain is clear. Alzheimer's diseasedamages and kills brain cells. A brain affected by Alzheimer's diseasehas many fewer cells and many fewer connections among surviving cellsthan does a healthy brain. As more and more brain cells die, Alzheimer'sleads to significant brain shrinkage. When doctors examine Alzheimer'sbrain tissue under the microscope, they see two types of abnormalitiesthat are considered hallmarks of the disease: plaques (clumps of aprotein called beta-amyloid that may damage and destroy brain cells inseveral ways, including interfering with cell-to-cell communication) andtangles (abnormal twists of tau proteins that lead to a failure in theinternal support and nutrient transport system upon which brain cellsdepend).

Liver

In some embodiments, provided peptides are used in the treatment ofinflammation associated with inflammatory conditions of the liver (e.g.hepatitis). Hepatitis is a medical condition defined by the inflammationof the liver and characterized by the presence of inflammatory cells inthe tissue of the organ. Hepatitis may occur with limited or nosymptoms, but often leads to jaundice, anorexia (poor appetite) andmalaise. Hepatitis is acute when it lasts less than six months andchronic when it persists longer. A group of viruses known as thehepatitis viruses cause most cases of hepatitis worldwide, but hepatitiscan also be caused by toxic substances (notably alcohol, certainmedications, some industrial organic solvents and plants), otherinfections and autoimmune diseases.

Initial features are of nonspecific flu-like symptoms, common to almostall acute viral infections and may include malaise, muscle and jointaches, fever, nausea or vomiting, diarrhea, and headache. More specificsymptoms, which can be present in acute hepatitis from any cause, are:profound loss of appetite, aversion to smoking among smokers, darkurine, yellowing of the eyes and skin (i.e., jaundice) and abdominaldiscomfort. Physical findings are usually minimal, apart from jaundicein a third and tender hepatomegaly (swelling of the liver) in about 10%.Some exhibit lymphadenopathy (enlarged lymph nodes) or splenomegaly(enlargement of the spleen). Acute viral hepatitis is more likely to beasymptomatic in younger people. Symptomatic individuals may presentafter convalescent stage of 7 to 10 days, with the total illness lasting2 to 6 weeks. A small proportion of people with acute hepatitis progressto acute liver failure, in which the liver is unable to clear harmfulsubstances from the circulation (leading to confusion and coma due tohepatic encephalopathy) and produce blood proteins (leading toperipheral edema and bleeding). This may become life-threatening andoccasionally requires a liver transplant.

Chronic hepatitis often leads to nonspecific symptoms such as malaise,tiredness and weakness, and often leads to no symptoms at all. It iscommonly identified on blood tests performed either for screening or toevaluate nonspecific symptoms. The occurrence of jaundice indicatesadvanced liver damage. On physical examination there may be enlargementof the liver. Extensive damage to and scarring of liver (i.e. cirrhosis)leads to weight loss, easy bruising and bleeding tendencies, peripheraledema (swelling of the legs) and accumulation of ascites (fluid in theperitoneal cavity). Eventually, cirrhosis may lead to variouscomplications: esophageal varices (enlarged veins in the wall of theesophagus that can cause life-threatening bleeding) hepaticencephalopathy (confusion and coma) and hepatorenal syndrome (kidneydysfunction). Acne, abnormal menstruation, lung scarring, inflammationof the thyroid gland and kidneys may be present in women with autoimmunehepatitis.

Kidney

In some embodiments, provided peptides are used in the treatment ofinflammation associated with inflammatory conditions of the kidney (e.g.nephritis). Nephritis is inflammation of the nephrons in the kidneys.Glomerulonephritis is inflammation of the glomeruli. (When the term“nephritis” is used without qualification, this is often the conditionmeant.) Glomeruli remove excess fluid, electrolytes and waste from thebloodstream and pass them into the urine. Also called glomerulardisease, glomerulonephritis can be acute—a sudden attack ofinflammation—or chronic—coming on gradually. Interstitial nephritis ortubulo-interstitial nephritis is inflammation of the spaces betweenrenal tubules.

If glomerulonephritis occurs on its own, it's known as primaryglomerulonephritis. If another disease, such as lupus or diabetes, isthe cause, it's called secondary glomerulonephritis. If severe orprolonged, the inflammation associated with glomerulonephritis candamage the kidneys. Signs and symptoms of glomerulonephritis may dependon the cause and whether one has the acute or chronic form. The firstindication that something is wrong may come from symptoms or from theresults of a routine urinalysis. Signs and symptoms may include: pink orcola-colored urine from red blood cells in the urine (hematuria), foamyurine due to excess protein (proteinuria), high blood pressure(hypertension), fluid retention (edema) with swelling evident in theface, hands, feet and abdomen, and fatigue from anemia or kidneyfailure. Glomerulonephritis can damage the kidneys so that they losetheir filtering ability. This can lead to the accumulation of dangerouslevels of fluid, electrolytes and waste in the body (called kidneyfailure) and deprive the bloodstream of necessary protein. Complicationsof glomerulonephritis may include: acute kidney failure, chronic kidneyfailure, high blood pressure and nephrotic syndrome.

Pancreas

In some embodiments, provided peptides are used in the treatment ofinflammation associated with inflammatory conditions of the pancreas(e.g. pancreatitis). Pancreatitis is inflammation in the pancreas. Thepancreas is a long, flat gland that sits tucked behind the stomach inthe upper abdomen. The pancreas produces enzymes that help digestion andhormones that help regulate the way the body processes sugar (glucose).Pancreatitis can occur as acute pancreatitis—meaning it appears suddenlyand lasts for days. Or pancreatitis can occur as chronic pancreatitis,which describes pancreatitis that occurs over many years. Mild cases ofpancreatitis may go away without treatment, but severe cases can causelife-threatening complications.

Signs and symptoms of pancreatitis may vary, depending on which type isexperienced. Acute pancreatitis signs and symptoms include: upperabdominal pain, abdominal pain that radiates to the back, abdominal painthat feels worse after eating, nausea, vomiting, and tenderness whentouching the abdomen. Chronic pancreatitis signs and symptoms include:upper abdominal pain, indigestion, losing weight without trying andoily, smelly stools (steatorrhea). Pancreatitis can cause seriouscomplications, including: breathing problems, diabetes, infection,kidney failure, malnutrition, pancreatic cancer, and pseudocyst.

Pancreatitis occurs when digestive enzymes produced in the pancreasbecome activated while inside the pancreas, causing damage to the organ.During normal digestion, the inactivated pancreatic enzymes move throughducts in the pancreas and travel to the small intestine, where theenzymes become activated and help with digestion. In pancreatitis, theenzymes become activated while still in the pancreas. This causes theenzymes to irritate the cells of the pancreas, causing inflammation andthe signs and symptoms associated with pancreatitis. With repeated boutsof acute pancreatitis, damage to the pancreas can occur and lead tochronic pancreatitis. Scar tissue may form in the pancreas, causing lossof function. A poorly functioning pancreas can cause digestion problemsand diabetes. A number of causes have been identified for acutepancreatitis and chronic pancreatitis, including: alcoholism,gallstones, abdominal surgery, certain medications, cigarette smoking,cystic fibrosis, endoscopic retrograde cholangiopancreatography (ERCP)(when used to treat gallstones), family history of pancreatitis, highcalcium levels in the blood (hypercalcemia), high levels of parathyroidhormone in the blood (hyperparathyroidism), high triglyceride levels inthe blood (hypertriglyceridemia), infection, injury to the abdomen, andpancreatic cancer.

Cardiovascular Disorders

In some embodiments, provided peptides are used in the treatment ofinflammation associated with cardiovascular disorders (e.g., cholesterolmetabolic disorders, oxygen free radical injury, ischemia, pulmonaryfibrosis, idiopathic pulmonary fibrosis, reperfusion injury, acutevaso-occlusive crisis in sickle cell anemia).

Pulmonary Fibrosis

In some embodiments, provided peptides are used in the treatment ofpulmonary fibrosis. Pulmonary fibrosis occurs when lung tissue becomesdamaged and scarred. This thickened, stiff tissue makes it moredifficult for the lungs to work properly. As pulmonary fibrosis worsens,a person become progressively more short of breath.

Signs and symptoms of pulmonary fibrosis include: shortness of breath(dyspnea), a dry cough, fatigue, unexplained weight loss, and achingmuscles and joints. The course of pulmonary fibrosis—and the severity ofsymptoms—can vary considerably from person to person. Some people becomeill very quickly with severe disease. Others have more moderate symptomsthat worsen over months or years. Complications of pulmonary fibrosismay include: high blood pressure in the lungs (pulmonary hypertension),right-sided heart failure (cor pulmonale), respiratory failure and lungcancer.

Pulmonary fibrosis scars and thickens the tissue around and between theair sacs (alveoli) in the lungs. This makes it more difficult for oxygento pass into the bloodstream. The damage can be caused by many differentthings—including airborne toxins in the workplace, certain lung diseasesand even some types of medical treatments. Long-term exposure to anumber of toxins and pollutants can damage the lungs. These may include:silica dust, asbestos fibers, grain dust, and bird and animal droppings.Some people who receive radiation therapy for lung or breast cancer showsigns of lung damage months or sometimes years after the initialtreatment. The severity of the damage depends on: how much of the lungwas exposed to radiation, the total amount of radiation administered,whether chemotherapy also was used, and the presence of underlying lungdisease. Many drugs can damage the lungs, especially: chemotherapydrugs, heart medications, and some antibiotics. Lung damage can alsoresult from: tuberculosis, pneumonia, systemic lupus erythematosus,rheumatoid arthritis, sarcoidosis, and scleroderma. The list ofsubstances and conditions that can lead to pulmonary fibrosis is long.Even so, in most cases, the cause is never found. Pulmonary fibrosiswith no known cause is called idiopathic pulmonary fibrosis. Researchershave several theories about what might trigger idiopathic pulmonaryfibrosis, including viruses and exposure to tobacco smoke. Additionally,because one type of idiopathic pulmonary fibrosis runs in families,heredity also is thought to play a role.

Reperfusion Injury

In some embodiments, provided peptides are used in the treatment ofreperfusion injury. Reperfusion injury is the tissue damage caused whenblood supply returns to the tissue after a period of ischemia or lack ofoxygen. The absence of oxygen and nutrients from blood during theischemic period creates a condition in which the restoration ofcirculation results in inflammation and oxidative damage through theinduction of oxidative stress rather than restoration of normalfunction.

The inflammatory response is partially responsible for the damage ofreperfusion injury. White blood cells, carried to the area by the newlyreturning blood, release a host of inflammatory factors such asinterleukins as well as free radicals in response to tissue damage.[1]The restored blood flow reintroduces oxygen within cells that damagescellular proteins, DNA, and the plasma membrane. Damage to the cell'smembrane may in turn cause the release of more free radicals. Suchreactive species may also act indirectly in redox signaling to turn onapoptosis. White blood cells may also bind to the endothelium of smallcapillaries, obstructing them and leading to more ischemia.

Reperfusion injury plays a part in the brain's ischemic cascade, whichis involved in stroke and brain trauma. Similar failure processes areinvolved in brain failure following reversal of cardiac arrest; controlof these processes is the subject of ongoing research. Repeated bouts ofischemia and reperfusion injury also are thought to be a factor leadingto the formation and failure to heal of chronic wounds such as pressuresores and diabetic foot ulcers. Continuous pressure limits blood supplyand causes ischemia, and the inflammation occurs during reperfusion. Asthis process is repeated, it eventually damages tissue enough to cause awound. In prolonged ischemia (60 minutes or more), hypoxanthine isformed as breakdown product of ATP metabolism. The enzyme xanthinedehydrogenase acts in reverse, that is as a xanthine oxidase as a resultof the higher availability of oxygen. This oxidation results inmolecular oxygen being converted into highly reactive superoxide andhydroxyl radicals. Xanthine oxidase also produces uric acid, which mayact as both a pro-oxidant and as a scavenger of reactive species such asperoxynitrite. Excessive nitric oxide produced during reperfusion reactswith superoxide to produce the potent reactive species peroxynitrite.Such radicals and reactive oxygen species attack cell membrane lipids,proteins, and glycosaminoglycans, causing further damage. They may alsoinitiate specific biological processes by redox signaling. Reperfusioninjury is a primary concern in liver transplantation surgery.

Acute Vaso-Occlusive Crisis in Sickle Cell Anemia

In some embodiments, provided peptides are used in the treatment ofpacute vaso-occlusive crisis. Sickle cell anemia is an inherited form ofanemia—a condition in which there aren't enough healthy red blood cellsto carry adequate oxygen throughout the body. Normally, red blood cellsare flexible and round, moving easily through blood vessels. In sicklecell anemia, the red blood cells become rigid and sticky and are shapedlike sickles or crescent moons. These irregularly shaped cells can getstuck in small blood vessels, which can slow or block blood flow andoxygen to parts of the body. This decreased or blocked circulation iscalled vaso-occlusive crisis.

The most common complaint is of pain, and recurrent episodes may causeirreversible organ damage. One of the most severe forms is the acutechest syndrome, which occurs as a result of infarction of the lungparenchyma; this infarction can rapidly result in death of the patientif not properly managed immediately.

The management of an acute event of vaso-occlusive crisis is the use ofpotent analgesic (opioids), rehydration with normal saline or ringerslactate, treatment of malaria (whether symptomatic or not) usingAtemisinin Combination Therapy, and use of adjunct therapy such asoxygen via face mask.

Chronic Obstructive Pulmonary Disease (COPD)

In some embodiments, provided peptides are used in the treatment ofChronic obstructive pulmonary disease (COPD). COPD refers to a group oflung diseases that block airflow and make breathing difficult. Emphysemaand chronic bronchitis are the two most common conditions that make upCOPD. Chronic bronchitis is an inflammation of the lining of thebronchial tubes, which carry air to and from the lungs. Emphysema occurswhen the air sacs (alveoli) at the end of the smallest air passages(bronchioles) in the lungs are gradually destroyed.

Symptoms of COPD often don't appear until significant lung damage hasoccurred, and they usually worsen over time. For chronic bronchitis, themain symptom is a cough that a person has at least three months a yearfor two consecutive years. Other signs and symptoms of COPD include:shortness of breath, especially during physical activities, wheezing,chest tightness, having to clear one's throat first thing in themorning, due to excess mucus in the lungs, a chronic cough that producessputum that may be clear, white, yellow or greenish, blueness of thelips or fingernail beds (cyanosis), frequent respiratory infections,lack of energy, and unintended weight loss (in later stages). Peoplewith COPD are also likely to experience episodes called exacerbations,during which their symptoms become worse and persist for days or longer.Complications of COPD include: respiratory infections, high bloodpressure, heart problems, lung cancer and depression.

The main cause of COPD is tobacco smoking. However, there are likelyother factors at play in the development of COPD, such as a geneticsusceptibility to the disease, because only about 20 percent of smokersdevelop COPD. Other irritants can cause COPD, including cigar smoke,secondhand smoke, pipe smoke, air pollution and workplace exposure todust, smoke or fumes. In the developing world, COPD often occurs inwomen exposed to fumes from burning fuel for cooking and heating inpoorly ventilated homes. In about 1 percent of people with COPD, thedisease results from a genetic disorder that causes low levels of aprotein called alpha-1-antitrypsin. Alpha-1-antitrypsin (AAt) is made inthe liver and secreted into the bloodstream to help protect the lungs.Alpha-1-antitrypsin deficiency can affect the liver as well as thelungs. Damage to the liver can occur in infants and children, not justadults with long smoking histories. For adults with COPD related to AAtdeficiency, treatment options are the same as those for people with morecommon types of COPD. Some people can be treated by replacing themissing AAt protein, which may prevent further damage to the lungs.

For patients whose COPD is linked to smoking, the most essential step inany treatment plan for COPD is to stop all smoking. There several kindsof medications to treat the symptoms and complications of COPD and theyinclude: bronchodilators, inhaled steroids, combination inhalers, oralsteroids, phosphodiesterase-4 inhibitors, theophylline and antibiotics.Additional therapies for people with moderate or severe COPD include:oxygen therapy and a pulmonary rehabilitation program. Surgery is anoption for some people with some forms of severe emphysema who aren'thelped sufficiently by medications alone. In lung volume reductionsurgery, the surgeon removes small wedges of damaged lung tissue. Thiscreates extra space in the chest cavity so that the remaining lungtissue and the diaphragm work more efficiently. Lung transplantation maybe an option for certain people who meet specific criteria.

Cystic Fibrosis

In some embodiments, provided peptides are used in the treatment ofcystic fibrosis. Cystic fibrosis is a life-threatening disorder thatcauses severe damage to the lungs and digestive system. An inheritedcondition, cystic fibrosis affects the cells that produce mucus, sweatand digestive juices. These secreted fluids are normally thin andslippery. But in cystic fibrosis, a defective gene causes the secretionsto become thick and sticky. Instead of acting as a lubricant, thesecretions plug up tubes, ducts and passageways, especially in the lungsand pancreas. Cystic fibrosis signs and symptoms vary, depending on theseverity of the disease. Even in the same person, symptoms may worsen orimprove as time passes. In some children, symptoms begin during infancy.Other people may not experience symptoms until adolescence or adulthood.Screening of newborns for cystic fibrosis is now performed in all 50states. As a result, the condition is diagnosed within the first monthof life, before symptoms develop. For people born before newbornscreening was performed, it's important to be aware of the signs andsymptoms of cystic fibrosis.

People with cystic fibrosis tend to have a higher than normal level ofsalt in their sweat. Parents often can taste the salt when they kisstheir children. Most of the other signs and symptoms of cystic fibrosisaffect the respiratory system or the digestive system. Thick and stickymucus clogging the passageways that carry air in and out of the lungslead to respiratory signs and symptoms including: a persistent coughthat produces thick spit (sputum) and mucus, wheezing, breathlessness, adecreased ability to exercise, repeated lung infections, and inflamednasal passages or a stuffy nose. The thick mucus can also block tubesthat carry digestive enzymes from the pancreas to the small intestine.Without those enzymes, the intestines can't fully absorb the nutrientsfrom food, resulting in digestive signs and symptoms that include:foul-smelling, greasy stools, poor weight gain and growth, intestinalblockage, particularly in newborns (meconium ileus), and severeconstipation. The sticky mucus associated with cystic fibrosis causesmany complications, the most common in the respiratory, digestive andreproductive systems. Respiratory system complications can include:bronchiectasis, chronic infections, nasal polyps, coughing up blood,pneumothorax, collapsed lung and respiratory failure. Digestive systemcomplications can include: nutritional deficiencies, diabetes, blockedbile duct, rectal prolapse and intussusception. Almost all men withcystic fibrosis are infertile because the tube that connects the testesand prostate gland (vas deferens) is either blocked with mucus ormissing entirely. Certain fertility treatments and surgical proceduressometimes make it possible for men with cystic fibrosis to becomefathers. Although women with cystic fibrosis may be less fertile thanother women, it's possible for them to conceive and to have successfulpregnancies. Other complications of cystic fibrosis include osteoporosisand electrolyte imbalances.

Acute Inflammatory Conditions

In some embodiments, provided peptides are used in the treatment ofinflammation associated with acute inflammatory conditions (e.g.,endotoxemia, sepsis and septicaemia, toxic shock syndrome and infectiousdisease (e.g., myocarditis, cardiomyopathy, acute endocarditis,pericarditis);

Acute Endocarditis

In some embodiments, provided peptides are used in the treatment ofacute endocarditis. Endocarditis is an infection of the inner lining ofthe heart (endocardium). Endocarditis typically occurs when bacteria orother germs from another part of the body, such as the mouth, spreadthrough the bloodstream and attach to damaged areas in the heart. Leftuntreated, endocarditis can damage or destroy heart valves and can leadto life-threatening complications. Endocarditis is uncommon in peoplewith healthy hearts. People at greatest risk of endocarditis have adamaged heart valve, an artificial heart valve or other heart defects.

Endocarditis may develop slowly or suddenly—depending on what's causingthe infection and whether the person has any underlying heart problems.The infection can infect different people differently, so signs andsymptoms vary. They may include: fever, chills, a new or changed heartmurmur, fatigue, aching joints and muscles, night sweats, shortness ofbreath, paleness, persistent cough, swelling of the feet, legs orabdomen, unexplained weight loss, blood in the urine, tenderness in thespleen, Osler's nodes and petechiae. Endocarditis can cause severalmajor complications, including: stroke and organ damage, infections inother parts of the body and heart failure.

Endocarditis occurs when germs enter the bloodstream, travel to theheart, and attach to abnormal heart valves or damaged heart tissue.Bacteria cause most cases, but fungi or other microorganisms also may beresponsible. Sometimes the culprit is one of many common bacteria thatlive in the mouth, throat or other parts of the body. The offendingorganism may enter the bloodstream through: everyday oral activities, aninfection of other medical condition, catheters or needles, or certaindental procedures. Typically, the immune system destroys bacteria thatmake it into the bloodstream. Even if bacteria reach the heart, they maypass through without causing an infection. Most people who developendocarditis have a diseased or damaged heart valve—an ideal spot forbacteria to settle. This damaged tissue in the endocardium providesbacteria with the roughened surface they need to attach and multiply.

Pericarditis

In some embodiments, provided peptides are used in the treatment ofpericarditis. Pericarditis is a swelling and irritation of thepericardium, the thin sac-like membrane that surrounds the heart.Pericarditis often causes chest pain and sometimes other symptoms.Pericarditis is usually sudden and short-lived (acute).

Acute pericarditis usually lasts less than a few weeks. Chronicpericarditis usually lasts six months or longer. In acute pericarditis,the most common symptom is sharp, stabbing chest pain behind thebreastbone or in the left side of the chest. However, some people withacute pericarditis describe their chest pain as dull, achy orpressure-like instead, and of varying intensity. The pain of acutepericarditis may travel into the left shoulder and neck. It oftenintensifies when one lies down or inhales deeply. Coughing, taking adeep breath or swallowing food also may make the pain worse. Sitting upand leaning forward can often ease the pain. At times, it may bedifficult to distinguish pericardial pain from the pain that occurs witha heart attack. Chronic pericarditis is usually associated with chronicinflammation and may result in fluid around the heart (pericardialeffusion). The most common symptom of chronic pericarditis is chestpain. Depending on the type, signs and symptoms of pericarditis mayinclude some or all of the following: sharp, piercing chest pain overthe center or left side of the chest, shortness of breath whenreclining, low-grade fever, an overall sense of weakness, fatigue orfeeling sick, dry cough, and abdominal or leg swelling. Many of thesymptoms of pericarditis are similar to those of other heart and lungconditions. For example, although the cause of acute chest pain may bepericarditis, the original cause could be a heart attack or a blood clotof the lungs (pulmonary embolus). Complications of pericarditis caninclude constrictive pericarditis (permanent thickening, scarring andcontraction of the pericardium) and cardiac tamponade (dramatic drop inblood pressure caused by excess fluid around the heart preventing theheart from filling and leaving the heart).

Adult Respiratory Distress Syndrome (ARDS)

In some embodiments, provided peptides are used in the treatment ofacute respiratory distress syndrome (ARDS). Acute respiratory distresssyndrome occurs when fluid builds up in the tiny, elastic air sacs(alveoli) in the lungs. More fluid in the lungs means less oxygen canreach the bloodstream. This deprives organs of the oxygen they need tofunction. Many people who develop ARDS don't survive. The risk of deathincreases with age and severity of illness. Of the people who do surviveARDS, some recover completely while others experience lasting damage totheir lungs.

The signs and symptoms of ARDS can vary in intensity, depending on itscause and severity. They include: severe shortness of breath, laboredand unusually rapid breathing, low blood pressure, and confusion andextreme tiredness. ARDS typically occurs in people who are alreadycritically ill or who have significant injuries. ARDS is extremelyserious, but thanks to improved treatments, more people are survivingit. However, many survivors end up with potentially serious—andsometimes lasting—complications, including: pulmonary fibrosis,collapsed lung (pneumothorax), blood clots, infections, abnormal lungfunction, and memory, cognitive and emotional problems.

The mechanical cause of ARDS is fluid leaked from the smallest bloodvessels in the lungs into the tiny air sacs where blood is oxygenated.Normally, a protective membrane keeps this fluid in the vessels. Severeillness or injury, however, can cause inflammation that undermines themembrane's integrity, leading to the fluid leakage of ARDS. The mostcommon underlying causes of ARDS include: sepsis, inhalation of harmfulsubstances, severe pneumonia and head or chest injury.

Scleroderma

In some embodiments, provided peptides are used in the treatment ofscleroderma. Scleroderma is a group of rare diseases that involve thehardening and tightening of the skin and connective tissue, the fibersthat provide the framework and support for the body. In some people,scleroderma affects only the skin. But in many people, scleroderma alsoharms structures beyond the skin, such as blood vessels, internal organsand the digestive tract. Signs and symptoms vary, depending on whichstructures are affected. Scleroderma affects women more often than menand most commonly occurs between the ages of 30 and 50. While there isno cure for scleroderma, a variety of treatments can ease symptoms andimprove quality of life.

Scleroderma's signs and symptoms vary, depending on which parts of thebody are involved. Nearly everyone who has scleroderma experiences ahardening and tightening of patches of skin. These patches may be shapedlike ovals or straight lines. The number, location and size of thepatches vary by type of scleroderma. Skin can appear shiny because it'sso tight, and movement of the affected area may be restricted. One ofthe earliest signs of scleroderma is an exaggerated response to coldtemperatures or emotional distress, which can cause numbness, pain orcolor changes in the fingers or toes. Called Raynaud's phenomenon, thiscondition also occurs in people who don't have scleroderma. In additionto acid reflux, which can damage the section of esophagus nearest thestomach, some people with scleroderma may also have problems absorbingnutrients if their intestinal muscles aren't moving food properlythrough the intestines. Rarely, scleroderma can affect the function ofthe heart, lungs or kidneys. These problems can become life-threatening.

Scleroderma complications range from mild to severe and can affect the:fingertips (the variety of Raynaud's phenomenon that occurs withscleroderma can be so severe that the restricted blood flow permanentlydamages the tissue at the fingertips, causing pits or skin sores(ulcers); in some cases, gangrene and amputation may follow), lungs(scarring of lung tissue (pulmonary fibrosis) can result in reduced lungfunction, reduced ability to breathe and reduced tolerance for exercise;patients can also develop high blood pressure in the arteries to thelungs (pulmonary hypertension)), kidneys (possibility of developingelevated blood pressure and an increased level of protein in the urine;more-serious effects of kidney complications may include renal crisis,which involves a sudden increase in blood pressure and rapid kidneyfailure), heart (scarring of heart tissue increases the risk of abnormalheartbeats (arrhythmias) and congestive heart failure, and can causeinflammation of the membranous sac surrounding the heart (pericarditis);scleroderma also can raise the pressure on the right side of the heartand cause it to wear out), teeth (severe tightening of facial skin cancause the mouth to become smaller and narrower, which may make it hardto brush the teeth or to even have them professionally cleaned; peoplewho have scleroderma often don't produce normal amounts of saliva, sothe risk of dental decay increases even more), digestive system(digestive problems associated with scleroderma can lead to acid refluxand difficulty swallowing—some describe feeling as if food gets stuckmidway down the esophagus—as well as bouts of constipation alternatingwith episodes of diarrhea) and sexual function (men who have sclerodermaoften experience erectile dysfunction; scleroderma may also affect thesexual function of women, by decreasing sexual lubrication andconstricting the vaginal opening).

Scleroderma results from an overproduction and accumulation of collagenin body tissues. Collagen is a fibrous type of protein that makes up thebody's connective tissues, including the skin. Although doctors aren'tsure what prompts this abnormal collagen production, the body's immunesystem appears to play a role. For unknown reasons, the immune systemturns against the body, producing inflammation and the overproduction ofcollagen.

Systemic Lupus Erythematosis

In some embodiments, provided peptides are used in the treatment ofsystemic lupus erythmatosis. Lupus is a chronic inflammatory diseasethat occurs when the body's immune system attacks its own tissues andorgans. Inflammation caused by lupus can affect many different bodysystems—including joints, skin, kidneys, blood cells, brain, heart andlungs. Lupus can be difficult to diagnose because its signs and symptomsoften mimic those of other ailments. The most distinctive sign oflupus—a facial rash that resembles the wings of a butterfly unfoldingacross both cheeks—occurs in many but not all cases of lupus.

Lupus occurs when the immune system attacks healthy tissue in the body.It's likely that lupus results from a combination of genetics andenvironment. It appears that people with an inherited predisposition forlupus may develop the disease when they come into contact with somethingin the environment that can trigger lupus. The cause for lupus in mostcases, however, is unknown. Some potential triggers include: sunlight(exposure to the sun may bring on lupus skin lesions or trigger aninternal response in susceptible people) and medications (lupus can betriggered by certain types of anti-seizure medications, blood pressuremedications and antibiotics; people who have drug-induced lupus usuallysee their symptoms go away when they stop taking the medication).

Inflammation caused by lupus can affect many areas of the body. Lupuscan cause serious kidney damage, and kidney failure is one of theleading causes of death among people with lupus. Signs and symptoms ofkidney problems may include generalized itching, chest pain, nausea,vomiting and leg swelling (edema). If the brain is affected by lupus, aperson may experience headaches, dizziness, behavior changes,hallucinations, and even strokes or seizures. Many people with lupusexperience memory problems and may have difficulty expressing theirthoughts. Lupus can lead to blood problems, including anemia andincreased risk of bleeding or blood clotting. It can also causeinflammation of the blood vessels (vasculitis). Having lupus increases aperson's chances of developing an inflammation of the chest cavitylining (pleurisy), which can make breathing painful. Lupus can causeinflammation of the heart muscle, arteries or heart membrane(pericarditis). The risk of cardiovascular disease and heart attacksincreases greatly as well. Lupus can also increase a person's risk ofinfection, cancer, bone tissue death (avascular necrosis) and pregnancycomplications).

Sjogren's Syndrome

In some embodiments, provided peptides are used in the treatment ofSjogren's syndrome. Sjogren's syndrome is a disorder of the immunesystem identified by its two most common symptoms—dry eyes and a drymouth. Sjogren's syndrome often accompanies other immune-systemdisorders, such as rheumatoid arthritis and lupus. In Sjogren'ssyndrome, the mucous membranes and moisture-secreting glands of the eyesand mouth are usually affected first—resulting in decreased productionof tears and saliva. Although a person can develop Sjogren's syndrome atany age, most people are older than 40 at the time of diagnosis. Thecondition is much more common in women.

The two main symptoms of Sjogren's syndrome are dry eyes and dry mouth.The eyes may burn, itch or feel gritty, as if there's sand in them andthe mouth may feel like it's full of cotton, making it difficult toswallow or speak. Some people with Sjogren's syndrome also experienceone of more of the following: joint pain, swelling and stiffness,swollen salivary glands—particularly the set located behind the jaw andin front of the ears, skin rashes or dry skin, vaginal dryness,persistent dry cough and prolonged fatigue.

Sjogren's syndrome is an autoimmune disorder. This means that the immunesystem mistakenly attacks the body's own cells and tissues. Scientistsaren't certain why some people develop Sjogren's syndrome and othersdon't. Certain genes put people at higher risk for the disorder, but itappears that a triggering mechanism—such as infection with a particularvirus or strain of bacteria—is also necessary. In Sjogren's syndrome,the immune system first targets the moisture-secreting glands of theeyes and mouth. But it can also damage other parts of the body, such asthe joints, thyroid, kidneys, liver, lungs, skin and nerves.

The most common complications of Sjogren's syndrome involve the eyes andmouth and include dental cavities, yeast infections of the mouth andvision problems. Less common complications may affect the lungs, kidneysor liver, unborn baby, lymph nodes or nerves.

Autoimmune Thyroiditis

In some embodiments, provided peptides are used in the treatment ofautoimmune thyroiditis. Autoimmune thyroiditis, is a disease in whichthe body interprets the thyroid glands and its hormone products T3, T4and TSH as threats, therefore producing special antibodies that targetthe thyroid's cells, thereby destroying it. It presents withhypothyroidism or hyperthyroidism and the presence or absence ofgoiters. Specialists clinically separate autoimmune thyroiditis into twocategories. If goiters are present, it is understood as Hashimoto'sThyroiditis. On the other hand, if the thyroid is atrophic, but does notpresent goiters, it is denominated Atrophic Thyroiditis. If the symptomsof thyroiditis appear in women after giving birth, it is attributed tosuch and therefore called Postpartum Thyroiditis. The effects of thisdisease are not permanent but transient. Symptoms may come and godepending on whether the patient receives treatment, and whether thetreatment is effective.

The symptoms for autoimmune thyroiditis may vary depending on whether itcauses hyperthyroidism or hypothyroidism. Hyperthyroidism can causesweating, rapid heart rate, anxiety, tremors, fatigue, difficultysleeping, sudden weight loss, and protruding eyes. Hypothyroidism cancause weight gain, fatigue, dry skin, hair loss, intolerance to cold,and constipation.

The exact cause for autoimmune thyroiditis is not known for certain, butresearchers have found high correlations with aspects such as genetics,high iodine consumption, and age. The disease is said to be inherited asa dominant trait because it has been reported that as many as fiftypercent of the first degree relatives of patients with some type ofautoimmune thyroiditis present thyroid antibodies in serum. Some studieshave even related it to chromosome 21 because of its high correlationwith patients with Down's syndrome and familial Alzheimer's disease.Still, this theory is doubted, since patients with Turner's syndromealso present a high prevalence of autoimmune thyroiditis (up to fiftypercent). Autoimmune thyroiditis has a higher prevalence in societiesthat have a higher intake of iodine in their diet, such as the UnitedStates and Japan. Also, the rate of lymphositic infiltration increasedin areas where the iodine intake was once low, but had been given iodinesupplementation. When in the presence of excess iodine, the gland isunable to avoid its inhibitory effect on the biosynthesis of thyroidhormones. This is why when a person has excess iodine in his or her dietor is administered with additional supplementation in areas wheresufficient iodine has been noted, it can induce reversiblehypothyroidism. The mean age of prevalence in women is higher than inmen by one year, 58 and 59 years old respectively. Autoimmunethyroiditis can also affect children. Although it is very rare forchildren under the age of five, it can occur, and it accounts for around40 percent of the cases in adolescents with goiters. In the case ofhypothyroidism, patients over the age of 45 have more chances ofdeveloping autoimmune thyroiditis.

Asthma

In some embodiments, provided peptides are used in the treatment ofasthma. Asthma is a condition in which the airways narrow and swell andproduce extra mucus. This can make breathing difficult and triggercoughing, wheezing and shortness of breath. For some people, asthma is aminor nuisance. For others, it can be a major problem that interfereswith daily activities and may lead to a life-threatening asthma attack.

Asthma symptoms range from minor to severe and vary from person toperson. A person may have infrequent asthma attacks, have symptoms onlyat certain times—such as when exercising—or have symptoms all the time.Asthma signs and symptoms include: shortness of breath, chest tightnessor pain, trouble sleeping caused by shortness of breath, coughing orwheezing, a whistling or wheezing sound when exhaling (wheezing is acommon sign of asthma in children), and coughing or wheezing attacksthat are worsened by a respiratory virus, such as a cold or the flu.Signs that a person's asthma is probably worsening include: asthma signsand symptoms that are more frequent and bothersome, increasingdifficulty breathing (measurable with a peak flow meter, a device usedto check how well one's lungs are working), and the need to use aquick-relief inhaler more often. For some people, asthma symptoms flareup in certain situations. In exercise-induced asthma, symptoms may beworse when the air is cold and dry. In occupational asthma, symptomsmays triggered by workplace irritants such as chemical fumes, gases ordust. In allergy-induced asthma, symptoms may be triggered by particularallergens, such as pet dander, cockroaches or pollen.

Cancer

In some embodiments, provided peptides are used in the treatment oftumors or cancers (e.g., soft tissue or solid tumors), such aspancreatic cancer, leukemia (e.g., Hodgkin's lymphoma), glioblastoma,astrocytoma, lymphoma, melanoma or prostate cancer.

Pancreatic Cancer

Pancreatic cancer begins in the tissues of the pancreas—an organ in theabdomen that lies horizontally behind the lower part of the stomach. Thepancreas secretes enzymes that aid digestion and hormones that helpregulate the metabolism of sugars. Pancreatic cancer often has a poorprognosis, even when diagnosed early. Pancreatic cancer typicallyspreads rapidly and is seldom detected in its early stages, which is amajor reason why it's a leading cause of cancer death. Signs andsymptoms may not appear until pancreatic cancer is quite advanced andsurgical removal isn't possible.

Signs and symptoms of pancreatic cancer often don't occur until thedisease is advanced. When signs and symptoms do appear, they mayinclude: upper abdominal pain that may radiate to the back, yellowing ofthe skin and the whites of the eyes (jaundice), loss of appetite, weightloss, depression, and blood clots. As pancreatic cancer progresses, itcan cause complications such as: jaundice, pain, bowel obstruction, andweight loss.

The cause of pancreatic cancer is unclear. Pancreatic cancer occurs whencells in the pancreas develop mutations in their DNA. These mutationscause cells to grow uncontrollably and to continue living after normalcells would die. These accumulating cells can form a tumor. Mostpancreatic cancer begins in the cells that line the ducts of thepancreas. This type of cancer is called pancreatic adenocarcinoma orpancreatic exocrine cancer. Rarely, cancer can form in thehormone-producing cells of the pancreas. This type of cancer is calledislet cell cancer or pancreatic endocrine cancer.

Melanoma

In some embodiments, provided peptides are used in the treatment ofinflammation associated with melanoma. Melanoma, the most serious typeof skin cancer, develops in the cells (melanocytes) that producemelanin—the pigment that gives skin its color. Melanoma can also form inthe eyes and, rarely, in internal organs, such as the intestines. Therisk of melanoma seems to be increasing in people under 40, especiallywomen. Knowing the warning signs of skin cancer can help ensure thatcancerous changes are detected and treated before the cancer has spread.Melanoma can be treated successfully if it is detected early.

Melanomas can develop anywhere on the body. They most often develop inareas that have had exposure to the sun, such as the back, legs, armsand face. Melanomas can also occur in areas that don't receive much sunexposure, such as the soles of the feet, palms of the hands andfingernail beds. These hidden melanomas are more common in people withdarker skin. The first melanoma signs and symptoms often are: a changein an existing mole and the development of a new pigmented orunusual-looking growth on the skin. Melanoma doesn't always begin as amole. It can also occur on otherwise normal-appearing skin.Characteristics of unusual moles that may indicate melanomas or otherskin cancers are: asymmetrical shape, irregular border, changes incolor, diameter, and evolving. Other suspicious changes in a mole mayinclude: scaliness, itching, spreading of pigment from the mole into thesurrounding skin, and oozing or bleeding. Cancerous (malignant) molesvary greatly in appearance. Some may show all of the changes listedabove, while others may have only one or two unusual characteristics.Melanomas can also develop in areas of the body that have little or noexposure to the sun, such as the spaces between toes and on the palms,soles, scalp or genitals. These are sometimes referred to as hiddenmelanomas, because they occur in places most people wouldn't think tocheck. When melanoma occurs in people with darker skin, it's more likelyto occur in a hidden area. Hidden melanomas include: melanoma under anail, melanoma in the mouth, digestive tract, urinary tract or vagina,and melanoma in the eye.

Melanoma occurs when something goes awry in the melanin-producing cells(melanocytes) that give skin its color. Normally, skin cells develop ina controlled and orderly way—healthy new cells push older cells towardthe skin's surface, where they die and eventually fall off. But whensome cells develop DNA damage, new cells may begin to grow out ofcontrol and can eventually form a mass of cancerous cells. Just whatdamages DNA in skin cells and how this leads to melanoma isn't clear.It's likely that a combination of factors, including environmental andgenetic factors, causes melanoma. Still, doctors believe exposure toultraviolet (UV) radiation from the sun and from tanning lamps and bedsis the leading cause of melanoma. UV light doesn't cause all melanomas,especially those that occur in places on the body that don't receiveexposure to sunlight. This indicates that other factors may contributeto the risk of melanoma.

Prostate Cancer

In some embodiments, provided peptides are used in the treatment ofinflammation associated with prostate cancer. Prostate cancer is cancerthat occurs in a man's prostate—a small walnut-shaped gland thatproduces the seminal fluid that nourishes and transports sperm. Prostatecancer is one of the most common types of cancer in men. Prostate cancerusually grows slowly and initially remains confined to the prostategland, where it may not cause serious harm. While some types of prostatecancer grow slowly and may need minimal or no treatment, other types areaggressive and can spread quickly. Prostate cancer that is detectedearly—when it's still confined to the prostate gland—has a better chanceof successful treatment. Prostate cancer may not cause signs or symptomsin its early stages. Prostate cancer that is more advanced may causesigns and symptoms such as: trouble urinating, decreased force in thestream of urine, blood in the urine, blood in the semen, general pain inthe lower back, hips or thighs, discomfort in the pelvic area, bonepain, and erectile dysfunction. Complications of prostate cancer and itstreatment include: metastasized cancer, incontinence and erectiledysfunction.

It's not clear what causes prostate cancer. Doctors know that prostatecancer begins when some cells in the prostate become abnormal. Mutationsin the abnormal cells' DNA cause the cells to grow and divide morerapidly than normal cells do. The abnormal cells continue living, whenother cells would die. The accumulating abnormal cells form a tumor thatcan grow to invade nearby tissue. Some abnormal cells can break off andspread (metastasize) to other parts of the body.

Transplant Rejection

In some embodiments, provided peptides are used in the treatment oftransplant rejection (e.g. destruction of pancreatic islet cells inislet cell transplantation, delayed graft failure in kidney or otherorgan transplantation).

Destruction of Pancreatic Islet Cells in Islet Cell Transplantation

In some embodiments, provided peptides are used in the treatment ofdestruction of pancreatic islet cells in islet cell transplantation. Thetwo types of pancreatic islet transplantation are allo-transplantationand auto-transplantation. Pancreatic islet allo-transplantation is aprocedure in which islets from the pancreas of a deceased organ donorare purified, processed, and transferred into another person. Pancreaticislet allo-transplantation is currently labeled an experimentalprocedure until the transplantation technology is considered successfulenough to be labeled therapeutic.

For each pancreatic islet allo-transplant infusion, researchers usespecialized enzymes to remove islets from the pancreas of a single,deceased donor. The islets are purified and counted in a lab. Transplantpatients typically receive two infusions with an average of 400,000 to500,000 islets per infusion. Once implanted, the beta cells in theseislets begin to make and release insulin. Pancreatic isletallo-transplantation is performed in certain patients with type 1diabetes whose blood glucose levels are difficult to control. The goalsof the transplant are to help these patients achieve normal bloodglucose levels with or without daily injections of insulin and to reduceor eliminate hypoglycemia unawareness—a dangerous condition in which aperson with diabetes cannot feel the symptoms of hypoglycemia, or lowblood glucose. When a person feels the symptoms of hypoglycemia, stepscan be taken to bring blood glucose levels back to normal.

Pancreatic islet allo-transplants are only performed at hospitals thathave received permission from the U.S. Food and Drug Administration(FDA) for clinical research on islet transplantation. The transplantsare often performed by a radiologist—a doctor who specializes in medicalimaging. The radiologist uses x rays and ultrasound to guide theplacement of a thin, flexible tube called a catheter through a smallincision in the upper abdomen—the area between the chest and hips—andinto the portal vein of the liver. The portal vein is the major veinthat supplies blood to the liver. The islets are then infused, orpushed, slowly into the liver through the catheter. Usually, the patientreceives a local anesthetic and a sedative. In some cases, a surgeonperforms the transplant using general anesthesia. Patients often needtwo or more transplants to get enough functioning islets to stop orreduce their need for insulin injections.

Pancreatic islet auto-transplantation is performed following totalpancreatectomy—the surgical removal of the whole pancreas—in patientswith severe and chronic, or long lasting, pancreatitis that cannot bemanaged by other treatments. This procedure is not consideredexperimental. Patients with type 1 diabetes cannot receive pancreaticislet auto-transplantation. The procedure is performed in a hospital,and the patient receives general anesthesia. The surgeon first removesthe pancreas and then extracts and purifies islets from the pancreas.Within hours, the islets are infused through a catheter into thepatient's liver. The goal is to give the body enough healthy islets tomake insulin.

As with any organ transplant, the recipient of an islet transplant musttake drugs every day to keep the body from rejecting the islets. Theimmune system is programmed to destroy bacteria, viruses, and tissue itrecognizes as “foreign,” including transplanted islets. In addition, theautoimmune response that destroyed transplant recipients' own islets inthe first place can recur and attack the transplanted islets.

Delayed Graft Failure in Kidney Transplantation

In some embodiments, provided peptides are used in the treatment ofdelayed graft failure in kidney transplantation. Delayed graft functionis a form of acute renal failure resulting in post-transplantationoliguria, increased allograft immunogenicity and risk of acute rejectionepisodes, and decreased long-term survival. Factors related to the donorand prerenal, renal, or postrenal transplant factors related to therecipient can contribute to this condition. Experimental studies havedemonstrated that both ischaemia and reinstitution of blood flow inischaemically damaged kidneys after hypothermic preservation activate acomplex sequence of events that sustain renal injury and play a pivotalpart in the development of delayed graft function. Elucidation of thepathophysiology of renal ischaemia and reperfusion injury hascontributed to the development of strategies to decrease the rate ofdelayed graft function, focusing on donor management, organ procurementand preservation techniques, recipient fluid management, andpharmacological agents (vasodilators, antioxidants, anti-inflammatoryagents). Several new drugs show promise in animal studies in preventingor ameliorating ischaemia-reperfusion injury and possibly delayed graftfunction, but definitive clinical trials are lacking. The goal ofmonotherapy for the prevention or treatment of is perhaps unattainable,and multidrug approaches or single drug targeting multiple signals willbe the next step to reduce post-transplantation injury and delayed graftfunction. Additional detail can be found in Perico et al, Lancet,364(9447), pages 1814-27 (2004), the entire disclosure of which ishereby incorporated by reference.

Stroke

In some embodiments, provided peptides are used in the treatment ofstroke. A stroke occurs when the blood supply to part of the brain isinterrupted or severely reduced, depriving brain tissue of oxygen andnutrients. Within minutes, brain cells begin to die.

It is important to note when signs and symptoms of a stroke beginbecause the length of time they have been present may guide treatmentdecisions. Signs and symptoms of stroke include: trouble with walking,trouble with speaking and understanding, paralysis or numbness of theface, arm or leg, trouble with seeing in one or both eyes, and headache.A stroke can sometimes cause temporary or permanent disabilities,depending on how long the brain suffers a lack of blood flow and whichpart was affected. Complications may include: paralysis or loss ofmuscle movement, difficulty talking or swallowing, memory loss orthinking difficulties, emotional problems, pain, and changes in behaviorand self-care.

A stroke occurs when the blood supply to the brain is interrupted orreduced. This deprives the brain of oxygen and nutrients, which cancause brain cells to die. A stroke may be caused by a blocked artery(ischemic stroke) or a leaking or burst blood vessel (hemorrhagicstroke). Some people may experience a temporary disruption of blood flowthrough their brain (transient ischemic attack). The majority of strokesare ischemic strokes. Ischemic strokes occur when the arteries to thebrain become narrowed or blocked, causing severely reduced blood flow(ischemia). The most common ischemic strokes include: thrombotic stroke(blood clot in one the arteries that supply blood to the brain) andembolic stroke (blood clot or other debris forms away from thebrain—commonly in the heart—and travels through the bloodstream to lodgein a narrow brain artery).

Hemorrhagic stroke occurs when a blood vessel in the brain leaks orruptures. Brain hemorrhages can result from many conditions that affectthe blood vessels, including uncontrolled high blood pressure(hypertension) and weak spots in blood vessel walls (aneurysms). A lesscommon cause of hemorrhage is the rupture of an arteriovenousmalformation (AVM)—an abnormal tangle of thin-walled blood vessels,present at birth. The types of hemorrhagic stroke include: intracerebralhemorrhage (blood vessel in brain bursts and spills into surroundingbrain tissue, damaging brain cells) and subarachnoid hemorrhage (arteryon or near the surface of the brain bursts and spills into the spacebetween the brain and the skull).

A transient ischemic attack (TIA)—also called a ministroke—is a briefepisode of symptoms similar to those one would have in a stroke. Atransient ischemic attack is caused by a temporary decrease in bloodsupply to part of the brain. TIAs often last less than five minutes.Like an ischemic stroke, a TIA occurs when a clot or debris blocks bloodflow to part of the brain. A TIA doesn't leave lasting symptoms becausethe blockage is temporary.

Neurodegenerative Diseases

In some embodiments, provided peptides are used in the treatment orprevention of neuronal death. In some embodiments, neuronal death isassociated with a neurodegenerative disease including, but not limitedto, amyotrophic lateral sclerosis (ALS), Huntington's disease,Alzheimer's disease, Parkinson's disease, multiple sclerosis, vascularcognitive impairment (VCI).

Amyotrophic Lateral Sclerosis (ALS)

In some embodiments, provided peptides are used in the treatment ofinflammation associated with amyotrophic lateral sclerosis (ALS). ALS isa serious neurological disease that causes muscle weakness, disabilityand eventually death. ALS is often called Lou Gehrig's disease, afterthe famous baseball player who was diagnosed with it in 1939. In theU.S., ALS and motor neuron disease (MND) are sometimes usedinterchangeably. Worldwide, ALS occurs in 1 to 3 people per 100,000. Inthe vast majority of cases—90 to 95 percent—doctors don't yet know whyALS occurs. About 5 to 10 percent of ALS cases are inherited.

Early signs and symptoms of ALS include: difficulty lifting the frontpart of the foot and toes (footdrop), weakness in the leg, feet orankles, hand weakness or clumsiness, slurring of speech or troubleswallowing, and muscle cramps and twitching in the arms, shoulders andtongue. The disease frequently begins in the hands, feet or limbs, andthen spreads to other parts of the body. As the disease advances, themuscles become progressively weaker until they're paralyzed. Iteventually affects chewing, swallowing, speaking and breathing.

In ALS, the nerve cells that control the movement of the musclesgradually die, so the muscles progressively weaken and begin to wasteaway. Up to 1 in 10 cases of ALS is inherited. But the remainder appearto occur randomly. Researchers are studying several possible causes ofALS, including: genetic mutations, chemical imbalances, disorganizedimmune response and mishandled proteins within neurons.

Huntington's Disease

In some embodiments, provided peptides are used in the treatment ofinflammation associated with Huntington's disease. Huntington's diseaseis an inherited disease that causes the progressive breakdown(degeneration) of nerve cells in the brain. Huntington's disease has abroad impact on a person's functional abilities and usually results inmovement, thinking (cognitive) and psychiatric disorders. Most peoplewith Huntington's disease develop signs and symptoms in their 40s or50s, but the onset of disease may be earlier or later in life. Whendisease onset begins before age 20, the condition is called juvenileHuntington's disease. Earlier onset often results in a somewhatdifferent presentation of symptoms and faster disease progression.

Huntington's disease usually causes movement, cognitive and psychiatricdisorders with a wide spectrum of signs and symptoms. Which symptomsappear first varies greatly among affected people. During the course ofthe disease, some disorders appear to be more dominant or have a greatereffect on functional ability. The movement disorders associated withHuntington's disease can include both involuntary movements andimpairments in voluntary movements: involuntary jerking or writhingmovements (chorea), involuntary, sustained contracture of muscles(dystonia), muscle rigidity, slow, uncoordinated fine movements, slow orabnormal eye movements, impaired gait, posture and balance, difficultywith the physical production of speech, and difficulty swallowingImpairments in voluntary movements—rather than the involuntarymovements—may have a greater impact on a person's ability to work,perform daily activities, communicate and remain independent.

Cognitive impairments often associated with Huntington's diseaseinclude: difficulty planning, organizing and prioritizing tasks,inability to start a task or conversation, lack of flexibility, or thetendency to get stuck on a thought, behavior or action (perseveration),lack of impulse control that can result in outbursts, acting withoutthinking and sexual promiscuity, problems with spatial perception thatcan result in falls, clumsiness or accidents, lack of awareness of one'sown behaviors and abilities, difficulty focusing on a task for longperiods, slowness in processing thoughts or “finding” words, anddifficulty in learning new information

The most common psychiatric disorder associated with Huntington'sdisease is depression. This isn't simply a reaction to receiving adiagnosis of Huntington's disease. Instead, depression appears to occurbecause of injury to the brain and subsequent changes in brain function.Signs and symptoms may include: feelings of sadness or unhappiness, lossof interest in normal activities, social withdrawal, insomnia orexcessive sleeping, fatigue, tiredness and loss of energy, feelings ofworthlessness or guilt, indecisiveness, distractibility and decreasedconcentration, frequent thoughts of death, dying or suicide, changes inappetite, and reduced sex drive. Other changes in mood or personality,but not necessarily specific psychiatric disorders, may include:irritability, apathy, anxiety, and sexual inhibition or inappropriatesexual behaviors.

The onset and progression of Huntington's disease in younger people maybe slightly different from that in adults. Problems that often presentthemselves early in the course of the disease include: loss ofpreviously learned academic or physical skills, rapid, significant dropin overall school performance, behavioral problems, contracted and rigidmuscles that affect gait (especially in young children), changes in finemotor skills that might be noticeable in skills such as handwriting,tremors or slight involuntary movements, and seizures.

After the onset of Huntington's disease, a person's functional abilitiesgradually worsen over time. The rate of disease progression and durationvaries. The time from disease onset to death is often about 10 to 30years. Juvenile onset usually results in death in fewer than 15 years.The clinical depression associated with Huntington's disease mayincrease the risk of suicide. Some research suggests that the greaterrisk of suicide occurs before a diagnosis is made and in middle stagesof the disease when a person has begun to lose independence. Eventually,a person with Huntington's disease requires help with all activities ofdaily living and care. Late in the disease, he or she will likely beconfined to a bed and unable to speak. However, a person's understandingof surroundings and interactions remain intact for a long time. Commoncauses of death include: pneumonia or other infections, injuries relatedto falls, and complications related to the inability to swallow.

Huntington's disease is caused by an inherited defect in a single gene.Huntington's disease is an autosomal dominant disorder, which means thata person needs only one copy of the defective gene to develop thedisorder. With the exception of genes on the sex chromosomes, a personinherits two copies of every gene—one copy from each parent. A parentwith a defective Huntington gene could pass along the defective copy ofthe gene or the healthy copy. Each child in the family, therefore, has a50 percent chance of inheriting the gene that causes the geneticdisorder.

Parkinson's Disease

In some embodiments, provided peptides are used in the treatment ofParkinson's disease. Parkinson's disease is a progressive disorder ofthe nervous system that affects movement. It develops gradually,sometimes starting with a barely noticeable tremor in just one hand. Butwhile tremor may be the most well-known sign of Parkinson's disease, thedisorder also commonly causes stiffness or slowing of movement.

Parkinson's disease symptoms and signs may vary from person to person.Early signs may be mild and may go unnoticed. Symptoms often begin onone side of the body and usually remain worse on that side, even aftersymptoms begin to affect both sides. Parkinson's signs and symptoms mayinclude: tremor, slowed movement (bradykinesia), rigid muscles, impairedposture and balance, loss of automatic movements, speech changes, andwriting changes. Parkinson's disease is often accompanied by theseadditional problems, which are variably treatable: thinkingdifficulties, depression and emotional changes, sleep problems anddisorders, bladder problems, constipation, and sexual dysfunction.

The cause of Parkinson's disease is unknown, but several factors appearto play a role, including: a person's genes and environmental triggers.Many changes occur in the brains of people with Parkinson's disease,including: clumps of specific substances called Lewy bodies within braincells, which are microscopic markers of Parkinson's disease, and aclumped form of the protein alpha-synuclein that is found in Lewy bodiesand can't be broken down by cells.

Vascular Cognitive Impairment (VCI)

In some embodiments, provided peptides are used in the treatment ofvascular cognitive impairment (VCI). Vascular cognitive impairment (VCI)is a spectrum of cognitive impairments caused by various types ofcerebrovascular disease that occurs as a result of interaction between avariety of vascular risk factors such as hypertension, obesity,dyslipidemia, diabetes mellitus, stroke and silent stroke. Included inthis spectrum is Vascular Dementia (VaD), (the second leading cause ofdementia after Alzheimer's disease (AD)) and Mild Vascular CognitiveImpairment (MVCI).

VCI may result from clinical stroke of the large vessels or frommicroangiopathic changes in the small cerebral vessels. Radiologicalfindings might include abnormally bright spots on a T2 weighted MRI scanin periventricular regions or in the deep white matter. This so-called“white matter disease” is commonly associated with vascular risk factorssuch as smoking and hypertension, and with subtle decline in cognitiveperformance with aging. Brain MRI might also show “lacunar infarcts”(spots which are hypointense on a T1 MRI scan) which are indicative ofsmall “silent strokes”, or hemorrhagic findings (small hemorrhagicfindings are often referred to as “microbleeds”). The relativeimportance and precise etiology of these findings remains a subject ofdebate. OCognitive domains commonly affected by VCI include psychomotorprocessing speed, executive function and verbal memory.

Traumatic Brain Injury (TBI)

In some embodiments, provided peptides are used in the treatment oftraumatic brain injury (TBI). TBI occurs when an external mechanicalforce causes brain dysfunction. Traumatic brain injury usually resultsfrom a violent blow or jolt to the head or body. An object penetratingthe skull, such as a bullet or shattered piece of skull, also can causetraumatic brain injury.

Traumatic brain injury can have wide-ranging physical and psychologicaleffects. Some signs or symptoms may appear immediately after thetraumatic event, while others may appear days or weeks later. The signsand symptoms of mild traumatic brain injury may include: loss ofconsciousness for a few seconds to a few minutes, no loss ofconsciousness, but a state of being dazed, confused or disoriented,memory or concentration problems, headache, dizziness or loss ofbalance, nausea or vomiting, sensory problems, such as blurred vision,ringing in the ears or a bad taste in the mouth, sensitivity to light orsound, mood changes or mood swings, feeling depressed or anxious,fatigue or drowsiness, difficulty sleeping, and sleeping more thanusual.

Moderate to severe traumatic brain injuries can include any of the signsand symptoms of mild injury, as well as the following symptoms that mayappear within the first hours to days after a head injury: loss ofconsciousness from several minutes to hours, profound confusion,agitation, combativeness or other unusual behavior, slurred speech,inability to awaken from sleep, weakness or numbness in fingers andtoes, loss of coordination, persistent headache or headache thatworsens, repeated vomiting or nausea, convulsions or seizures, dilationof one or both pupils of the eyes, and clear fluids draining from thenose or ears.

Infants and young children with brain injuries may lack thecommunication skills to report headaches, sensory problems, confusionand similar symptoms. In a child with traumatic brain injury, thefollowing signs and symptoms may be observed: change in eating ornursing habits, persistent crying and inability to be consoled, unusualor easy irritability, change in ability to pay attention, change insleep habits, sad or depressed mood, and loss of interest in favoritetoys or activities.

Several complications can occur immediately or soon after a traumaticbrain injury. Severe injuries increase the risk of a greater number ofcomplications and more-severe complications. Complications associatedwith TBI include: altered consciousness (including coma, vegetativestate, minimally conscious state, and locked-in syndrome). Some peoplewith TBI will have seizures within the first week. Some serious injuriesmay result in recurring seizures, called post-traumatic epilepsy.Additionally, cerebrospinal fluid may build up in the spaces in thebrain (cerebral ventricles) of some people who have had traumatic braininjuries, causing swelling and increased pressure in the brain. Skullfractures or penetrating wounds can tear the layers of protectivetissues (meninges) that surround the brain. This can enable bacteria toenter the brain and cause infections. An infection of the meninges(meningitis) could spread to the rest of the nervous system if nottreated. Several small or large blood vessels in the brain may bedamaged in a traumatic brain injury. This damage could lead to a stroke,blood clots or other problems. Injuries to the base of the skull candamage nerves that emerge directly from the brain (cranial nerves).Cranial nerve damage may result in: paralysis of facial muscles, damageto the nerves responsible for eye movements, which can cause doublevision, damage to the nerves that provide sense of smell, loss ofvision, loss of facial sensation, and swallowing problems.

Most people who have had a significant brain injury will experiencechanges in their cognitive skills. TBI can result in problems with manyskills, including: memory, learning, reasoning, problem solving, speedof mental processing, judgment, attention or concentration,multitasking, organization, decision making, beginning or completingtasks, and communication problems. Language and communications problemsare common following TBI. These problems can cause frustration, conflictand misunderstanding for people with a TBI, as well as family members,friends and care providers. Communication problems may include:difficulty understanding speech or writing, difficulty speaking orwriting, difficulty deciphering nonverbal signals, inability to organizethoughts and ideas, inability to use the muscles needed to form words(dysarthria), problems with changes in tone, pitch or emphasis toexpress emotions, attitudes or subtle differences in meaning, troublestarting or stopping conversations, trouble with turn taking or topicselection, trouble reading cues from listeners, and trouble followingconversations.

People who've experienced brain injury often experience changes inbehaviors. These may include: difficulty with self-control, lack ofawareness of abilities, risky behavior, inaccurate self-image,difficulty in social situations, and verbal or physical outbursts.Emotional changes may include: depression, anxiety, mood swings,irritability, lack of empathy for others, anger, insomnia, and changesin self-esteem. Problems involving senses may include: persistentringing in the ears, difficulty recognizing objects, impaired hand-eyecoordination, blind spots or double vision, a bitter taste or a badsmell, skin tingling, pain or itching, and trouble with balance ordizziness. A TBI may increase the risk of diseases that result in thegradual degeneration of brain cells and gradual loss of brain functions.These include: Alzheimer's disease, Parkinson's disease, and dementiapugilistica.

Traumatic brain injury is caused by a blow or other traumatic injury tothe head or body. The degree of damage can depend on several factors,including the nature of the event and the force of impact. Injury mayinclude one or more of the following factors: damage to brain cells maybe limited to the area directly below the point of impact on the skull,a severe blow or jolt can cause multiple points of damage because thebrain may move back and forth in the skull, a severe rotational orspinning jolt can cause the tearing of cellular structures, a blast, asfrom an explosive device, can cause widespread damage, an objectpenetrating the skull can cause severe, irreparable damage to braincells, blood vessels and protective tissues around the brain, andbleeding in or around the brain, swelling, and blood clots can disruptthe oxygen supply to the brain and cause wider damage. Common eventscausing traumatic brain injury include the following: falls,vehicle-related collisions, violence, sports injuries and explosiveblasts and other combat injuries.

Pharmaceutical Compositions

In accordance with the methods of the invention, provided peptides asdescribed herein can be administered to a subject alone (e.g., as apurified peptide or compound), or as a component of a composition ormedicament (e.g., in the manufacture of a medicament for the treatmentof the disease), as described herein. The compositions can be formulatedwith a physiologically acceptable carrier or excipient to prepare apharmaceutical composition. The carrier and composition can be sterile.The formulation should suit the mode of administration, for exampleintravenous or subcutaneous administration. Methods of formulatingcompositions are known in the art (see, e.g., Remington'sPharmaceuticals Sciences, 17th Edition, Mack Publishing Co., (Alfonso R.Gennaro, editor) (1989)). In some embodiments, a composition maycomprise one or more liposomes, nanoparticles, and/or sterile aqueous ornon-aqueous solutions, suspensions, and emulsions.

Suitable pharmaceutically acceptable carriers include but are notlimited to water, salt solutions (e.g., NaCl), saline, buffered saline,alcohols, glycerol, ethanol, gum arabic, vegetable oils, benzylalcohols, polyethylene glycols, gelatin, carbohydrates such as lactose,amylose or starch, sugars such as mannitol, sucrose, or others,dextrose, magnesium stearate, talc, silicic acid, viscous paraffin,perfume oil, fatty acid esters, hydroxymethylcellulose, polyvinylpyrolidone, etc., as well as combinations thereof. The pharmaceuticalpreparations can, if desired, be mixed with auxiliary agents (e.g.,lubricants, preservatives, stabilizers, wetting agents, emulsifiers,salts for influencing osmotic pressure, buffers, coloring and/oraromatic substances and the like) which do not deleteriously react withthe active compounds or interference with their activity. In a preferredembodiment, a water-soluble carrier suitable for intravenousadministration is used.

The composition or medicament, if desired, can also contain minoramounts of wetting or emulsifying agents, or pH buffering agents. Acomposition can be a liquid solution, suspension, emulsion, tablet,pill, capsule, sustained release formulation, or powder. The compositioncan also be formulated as a suppository, with traditional binders andcarriers such as triglycerides. Oral formulations can include standardcarriers such as pharmaceutical grades of mannitol, lactose, starch,magnesium stearate, polyvinyl pyrollidone, sodium saccharine, cellulose,magnesium carbonate, etc.

The composition or medicament can be formulated in accordance with theroutine procedures as a pharmaceutical composition adapted foradministration to human beings. For example, in a preferred embodiment,a composition for intravenous administration typically is a solution insterile isotonic aqueous buffer. Where necessary, the composition mayalso include a solubilizing agent and a local anesthetic to ease pain atthe site of the injection. Generally, the ingredients are suppliedeither separately or mixed together in unit dosage form, for example, asa dry lyophilized powder or water free concentrate in a hermeticallysealed container such as an ampule or sachette indicating the quantityof active agent. Where the composition is to be administered byinfusion, it can be dispensed with an infusion bottle containing sterilepharmaceutical grade water, saline or dextrose/water. Where thecomposition is administered by injection, an ampule of sterile water forinjection or saline can be provided so that the ingredients may be mixedprior to administration.

Provided peptides as described herein can be formulated as neutral orsalt forms. Pharmaceutically acceptable salts include those formed withfree amino groups such as those derived from hydrochloric, phosphoric,acetic, oxalic, tartaric acids, etc., and those formed with freecarboxyl groups such as those derived from sodium, potassium, ammonium,calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

Dosing

In some embodiments, a provided peptides and/or compositions areadministered in a therapeutically effective amount and/or according to adosing regimen that is correlated with a particular desired outcome(e.g., with treating or reducing risk for disease).

Any provided peptide as described herein (or a composition or medicamentcontaining provided peptides as described herein) may be administered byany appropriate route. In some embodiments, provided peptides asdescribed herein is administered intravenously. In some embodiments,provided peptides as described herein is administered subcutaneously. Asused herein, the term “subcutaneous tissue” is defined as a layer ofloose, irregular connective tissue immediately beneath the skin. Forexample, the subcutaneous administration may be performed by injecting acomposition into areas including, but not limited to, thigh region,abdominal region, gluteal region, or scapular region. In otherembodiments, provided peptides as described herein is administered bydirect administration to a target tissue, such as heart or muscle (e.g.,intramuscular), tumor (intratumorally), nervous system (e.g., directinjection into the brain; intraventricularly; intrathecally), or othertarget tissue such as the liver, kidney, etc. Alternatively, providedpeptides as described herein (or a composition or medicament containinga peptide described herein) can be administered via inhalation,intraperitoneally, parenterally, intradermally, transdermally, ortransmucosally (e.g., orally or nasally). More than one route can beused concurrently, if desired.

In some embodiments, provided peptides and/or compositions areadministered in a therapeutically effective amount and/or according to adosing regimen that is correlated with a particular desired outcome(e.g., with treating or reducing risk for disease).

Particular doses or amounts to be administered in accordance with thepresent invention may vary, for example, depending on the nature and/orextent of the desired outcome, on particulars of route and/or timing ofadministration, and/or on one or more characteristics (e.g., weight,age, personal history, genetic characteristic, lifestyle parameter,severity of cardiac defect and/or level of risk of cardiac defect, etc.,or combinations thereof). Such doses or amounts can be determined bythose of ordinary skill. In some embodiments, an appropriate dose oramount is determined in accordance with standard clinical techniques.For example, in some embodiments, an appropriate dose or amount is adose or amount sufficient to reduce one or more symptoms by 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35,40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100% or more.Alternatively or additionally, in some embodiments, an appropriate doseor amount is determined through use of one or more in vitro or in vivoassays to help identify desirable or optimal dosage ranges or amounts tobe administered.

In various embodiments, provided peptides, or a composition comprisingsuch peptides, is administered at a therapeutically effective amount. Asused herein, the term “therapeutically effective amount” or“therapeutically effective dosage amount” is largely determined based onthe total amount of the therapeutic agent contained in thepharmaceutical compositions of the present invention. Generally, atherapeutically effective amount is sufficient to achieve a meaningfulbenefit to the subject (e.g., treating, modulating, curing, preventingand/or ameliorating the underlying disease or condition). In someparticular embodiments, appropriate doses or amounts to be administeredmay be extrapolated from dose-response curves derived from in vitro oranimal model test systems.

Therapeutically effective dosage amounts of provided peptides, includingderivatives, analogs, and/or salts thereof, may be present in varyingamounts in various embodiments. In some embodiments, a therapeuticallyeffective dosage amount can be, for example, about 1-10,000 μg/kg, about5-1,500 μg/kg, about 100-1,000 μg/kg, or 50-500 μg/kg. In someembodiments, the therapeutically effective dosage amount can be, forexample, about 1 μg/kg, 2.5 μg/kg, 5 μg/kg, 10 μg/kg, 20 μg/kg, 30μg/kg, 40 μg/kg, 50 μg/kg, 60 mg/kg, 70 μg/kg, 80 μg/kg, 90 μg/kg, 100μg/kg, 150 μg/kg, 200 μg/kg, 250 μg/kg, 300 μg/kg, 400 μg/kg, 500 μg/kg,600 μg/kg, 700 μg/kg, 800 μg/kg, 900 μg/kg, 1000 μg/kg, or 1500 μg/kg.The effective dose for a particular individual can be varied (e.g.,increased or decreased) over time, depending on the needs of theindividual. In some embodiments, the therapeutically effective amountdescribed herein is provided in one dose. In some embodiments, thetherapeutically effective amount described herein is provided in oneday.

In other embodiments, a therapeutically effective dosage amount may be,for example, about 0.001 mg/kg weight to 500 mg/kg weight, e.g., fromabout 0.001 mg/kg weight to 400 mg/kg weight, from about 0.001 mg/kgweight to 300 mg/kg weight, from about 0.001 mg/kg weight to 200 mg/kgweight, from about 0.001 mg/kg weight to 100 mg/kg weight, from about0.001 mg/kg weight to 90 mg/kg weight, from about 0.001 mg/kg weight to80 mg/kg weight, from about 0.001 mg/kg weight to 70 mg/kg weight, fromabout 0.001 mg/kg weight to 60 mg/kg weight, from about 0.001 mg/kgweight to 50 mg/kg weight, from about 0.001 mg/kg weight to 40 mg/kgweight, from about 0.001 mg/kg weight to 30 mg/kg weight, from about0.001 mg/kg weight to 25 mg/kg weight, from about 0.001 mg/kg weight to20 mg/kg weight, from about 0.001 mg/kg weight to 15 mg/kg weight, fromabout 0.001 mg/kg weight to 10 mg/kg weight. In some embodiments, thetherapeutically effective amount described herein is provided in onedose. In some embodiments, the therapeutically effective amountdescribed herein is provided in one day.

In still other embodiments, a therapeutically effective dosage amountmay be, for example, about 0.001 mg/kg weight to about 1 mg/kg weight,e.g. from about 0.001 mg/kg weight to about 0.9 mg/kg weight, from about0.001 mg/kg weight to about 0.8 mg/kg weight, from about 0.001 mg/kgweight to about 0.8 mg/kg weight, from about 0.001 mg/kg weight to about0.7 mg/kg weight, from about 0.001 mg/kg weight to about 0.6 mg/kgweight, from about 0.001 mg/kg weight to about 0.5 mg/kg weight, fromabout 0.01 mg/kg weight to about 1 mg/kg weight, from about 0.01 mg/kgweight to about 0.9 mg/kg weight, from about 0.01 mg/kg weight to about0.8 mg/kg weight, from about 0.01 mg/kg weight to about 0.7 mg/kgweight, from about 0.01 mg/kg weight to about 0.6 mg/kg weight, fromabout 0.01 mg/kg weight to about 0.5 mg/kg weight, from about 0.02 mg/kgweight to about 1 mg/kg weight, from about 0.02 mg/kg weight to about0.9 mg/kg weight, from about 0.02 mg/kg weight to about 0.8 mg/kgweight, from about 0.02 mg/kg weight to about 0.7 mg/kg weight, fromabout 0.02 mg/kg weight to about 0.6 mg/kg weight, from about 0.02 mg/kgweight to about 0.5 mg/kg weight, from about 0.03 mg/kg weight to about1 mg/kg weight, from about 0.03 mg/kg weight to about 0.9 mg/kg weight,from about 0.03 mg/kg weight to about 0.8 mg/kg weight, from about 0.03mg/kg weight to about 0.7 mg/kg weight, from about 0.03 mg/kg weight toabout 0.6 mg/kg weight, from about 0.03 mg/kg weight to about 0.5 mg/kgweight, from about 0.04 mg/kg weight to about 1 mg/kg weight, from about0.04 mg/kg weight to about 0.9 mg/kg weight, from about 0.04 mg/kgweight to about 0.8 mg/kg weight, from about 0.04 mg/kg weight to about0.7 mg/kg weight, from about 0.04 mg/kg weight to about 0.6 mg/kgweight, from about 0.04 mg/kg weight to about 0.5 mg/kg weight, fromabout 0.05 mg/kg weight to about 1 mg/kg weight, from about 0.05 mg/kgweight to about 0.9 mg/kg weight, from about 0.05 mg/kg weight to about0.8 mg/kg weight, from about 0.05 mg/kg weight to about 0.7 mg/kgweight, from about 0.05 mg/kg weight to about 0.6 mg/kg weight, fromabout 0.05 mg/kg weight to about 0.5 mg/kg weight. In some embodiments,the therapeutically effective amount described herein is provided in onedose. In some embodiments, the therapeutically effective amountdescribed herein is provided in one day.

In still other embodiments, a therapeutically effective dosage amountmay be, for example, about 0.0001 mg/kg weight to 0.1 mg/kg weight, e.g.from about 0.0001 mg/kg weight to 0.09 mg/kg weight, from about 0.0001mg/kg weight to 0.08 mg/kg weight, from about 0.0001 mg/kg weight to0.07 mg/kg weight, from about 0.0001 mg/kg weight to 0.06 mg/kg weight,from about 0.0001 mg/kg weight to 0.05 mg/kg weight, from about 0.0001mg/kg weight to about 0.04 mg/kg weight, from about 0.0001 mg/kg weightto 0.03 mg/kg weight, from about 0.0001 mg/kg weight to 0.02 mg/kgweight, from about 0.0001 mg/kg weight to 0.019 mg/kg weight, from about0.0001 mg/kg weight to 0.018 mg/kg weight, from about 0.0001 mg/kgweight to 0.017 mg/kg weight, from about 0.0001 mg/kg weight to 0.016mg/kg weight, from about 0.0001 mg/kg weight to 0.015 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.014 mg/kg weight, from about 0.0001 mg/kgweight to 0.013 mg/kg weight, from about 0.0001 mg/kg weight to 0.012mg/kg weight, from about 0.0001 mg/kg weight to 0.011 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.01 mg/kg weight, from about 0.0001 mg/kgweight to 0.009 mg/kg weight, from about 0.0001 mg/kg weight to 0.008mg/kg weight, from about 0.0001 mg/kg weight to 0.007 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.006 mg/kg weight, from about 0.0001 mg/kgweight to 0.005 mg/kg weight, from about 0.0001 mg/kg weight to 0.004mg/kg weight, from about 0.0001 mg/kg weight to 0.003 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.002 mg/kg weight. In some embodiments,the therapeutically effective dose may be 0.0001 mg/kg weight, 0.0002mg/kg weight, 0.0003 mg/kg weight, 0.0004 mg/kg weight, 0.0005 mg/kgweight, 0.0006 mg/kg weight, 0.0007 mg/kg weight, 0.0008 mg/kg weight,0.0009 mg/kg weight, 0.001 mg/kg weight, 0.002 mg/kg weight, 0.003 mg/kgweight, 0.004 mg/kg weight, 0.005 mg/kg weight, 0.006 mg/kg weight,0.007 mg/kg weight, 0.008 mg/kg weight, 0.009 mg/kg weight, 0.01 mg/kgweight, 0.02 mg/kg weight, 0.03 mg/kg weight, 0.04 mg/kg weight, 0.05mg/kg weight, 0.06 mg/kg weight, 0.07 mg/kg weight, 0.08 mg/kg weight,0.09 mg/kg weight, or 0.1 mg/kg weight. The effective dose for aparticular individual can be varied (e.g., increased or decreased) overtime, depending on the needs of the individual. In some embodiments, thetherapeutically effective amount described herein is provided in onedose. In some embodiments, the therapeutically effective amountdescribed herein is provided in one day.

In some embodiments, the provided peptides, and/or a compositioncomprising such peptides, is administered at an effective dose rangingfrom about 1-1,000 μg/kg/day (e.g., ranging from about 1-900 μg/kg/day,1-800 μg/kg/day, 1-700 μg/kg/day, 1-600 μg/kg/day, 1-500 μg/kg/day,1-400 μg/kg/day, 1-300 μg/kg/day, 1-200 μg/kg/day, 1-100 μg/kg/day, 1-90μg/kg/day, 1-80 μg/kg/day, 1-70 μg/kg/day, 1-60 μg/kg/day, 1-50μg/kg/day, 1-40 μg/kg/day, 1-30 μg/kg/day, 1-20 μg/kg/day, 1-10μg/kg/day). In some embodiments, the provided peptides, and/or acomposition comprising such peptides, are administered at an effectivedose ranging from about 1-500 μg/kg/day. In some embodiments, theprovided peptides, and/or a composition comprising such peptides, areadministered at an effective dose ranging from about 50-500 μg/kg/day.In some embodiments, the provided peptides, and/or a compositioncomprising such peptides, are administered at an effective dose rangingfrom about 1-100 μg/kg/day. In some embodiments, the provided peptides,and/or a composition comprising such peptides, are administered at aneffective dose ranging from about 1-60 μg/kg/day In some embodiments,the provided peptides, and/or a composition comprising such peptides,are administered at an effective dose selected from about 1, 2, 4, 6, 8,10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350,400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1,000μg/kg/day. The effective dose for a particular individual can be varied(e.g., increased or decreased) over time, depending on the needs of theindividual. In some embodiments, the therapeutically effective amountdescribed herein is provided in one dose. In some embodiments, thetherapeutically effective amount described herein is provided in oneday.

In some embodiments, In some embodiments, the provided peptides, and/ora composition comprising such peptides, are administered at an effectivedose from about 1-1,000 pg/kg/day (e.g., ranging from about 1-900pg/kg/day, 1-800 pg/kg/day, 1-700 pg/kg/day, 1-600 pg/kg/day, 1-500pg/kg/day, 1-400 pg/kg/day, 1-300 pg/kg/day, 1-200 pg/kg/day, 1-100pg/kg/day, 1-90 pg/kg/day, 1-80 pg/kg/day, 1-70 pg/kg/day, 1-60pg/kg/day, 1-50 pg/kg/day, 1-40 pg/kg/day, 1-30 pg/kg/day, 1-20pg/kg/day, 1-10 pg/kg/day). In some embodiments, the provided peptides,and/or a composition comprising such peptides, are administered at aneffective dose ranging from about 1-60 μg/kg/day. In some embodiments,the provided peptides, and/or a composition comprising such peptides,are administered at an effective dose selected from about 1, 2, 4, 6, 8,10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350,400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1,000pg/kg/day. The effective dose for a particular individual can be varied(e.g., increased or decreased) over time, depending on the needs of theindividual. In some embodiments, the therapeutically effective amountdescribed herein is provided in one dose. In some embodiments, thetherapeutically effective amount described herein is provided in oneday.

In some embodiments, a composition is provided as a pharmaceuticalformulation. In some embodiments, a pharmaceutical formulation is orcomprises a unit dose amount for administration in accordance with adosing regimen correlated with achievement of the reduced incidence orrisk of disease.

In some embodiments, a formulation comprising provided peptides asdescribed herein is administered as a single dose. In some embodiments,a formulation comprising provided peptides as described herein isadministered at regular intervals. Administration at an “interval,” asused herein, indicates that the therapeutically effective amount isadministered periodically (as distinguished from a one-time dose). Theinterval can be determined by standard clinical techniques. In someembodiments, a formulation comprising provided peptides as describedherein is administered bimonthly, monthly, twice monthly, triweekly,biweekly, weekly, twice weekly, thrice weekly, daily, twice daily, orevery six hours. The administration interval for a single individualneed not be a fixed interval, but can be varied over time, depending onthe needs of the individual.

As used herein, the term “bimonthly” means administration once per twomonths (i.e., once every two months); the term “monthly” meansadministration once per month; the term “triweekly” means administrationonce per three weeks (i.e., once every three weeks); the term “biweekly”means administration once per two weeks (i.e., once every two weeks);the term “weekly” means administration once per week; and the term“daily” means administration once per day.

In some embodiments, a formulation comprising provided peptides asdescribed herein is administered at regular intervals indefinitely. Insome embodiments, a formulation comprising provided peptides asdescribed herein is administered at regular intervals for a definedperiod. In some embodiments, a formulation comprising provided peptidesas described herein is administered at regular intervals for 5 years, 4,years, 3, years, 2, years, 1 year, 11 months, 10 months, 9 months, 8months, 7 months, 6 months, 5 months, 4 months, 3 months, 2 months, amonth, 3 weeks, 2, weeks, a week, 6 days, 5 days, 4 days, 3 days, 2 daysor a day.

Combination Therapies

In some embodiments, the provided peptides, and/or a compositioncomprising such peptides, may be administered in combination with one ormore known therapeutic agents currently used for prophylaxis andtreatment of diseases, disorders and conditions in which a CXCR mediatedpathway is implicated (e.g. inflammation, stroke, traumatic braininjury, pancreatic cancer, and neurodegenerative diseases). These agentsinclude anti-inflammatory agents, anticancer agents, neuroprotectiveagents, thrombolytic agents, immunosuppressants, antioxidants, α- orβ-adrenergic agonist or antagonist or a NMDA receptor antagonist, allwell known in the art.

Anti-inflammatory drugs or agents which may be coadministered withcompounds according to the present invention include at least one agentsuch as a non-steroidal anti-inflammatory drug (NSAID), for exampleaspirin, ibuprofen, naproxen, an IMSAID such as PHE-GLU-GLY (FEG) orD-isomeric feG, a bioactive compound such as plumbagin or plumericin, orany one or more supplement including hyssop, ginger, tumeric, willowbark, hyperforin, omega-3 fatty acids and ginger, among others.

Anticancer agents which may be used in combination with compoundsaccording to the present invention include, for example,microtubule-stabilizing agents, microtubule-disruptor agents, alkylatingagents, antimetabolites, epidophyllotoxins, antineoplastic enzymes,topoisomerase inhibitors, inhibitors of cell cycle progression, andplatinum coordination complexes, among others. These may be selectedfrom the group consisting of everolimus, trabectedin, abraxane, TLK 286,AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244(ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin,vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, aFLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurorakinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HDACinhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFRTK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3 kinaseinhibitors, an AKT inhibitor, a JAK/STAT inhibitor, a checkpoint-1 or 2inhibitor, a focal adhesion kinase inhibitor, a Map kinase kinase (mek)inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib,nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu,nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin,tetrandrine, rubitecan, tesmilifene, oblimersen, ticilimumab,ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490,cilengitide, gimatecan, IL13-PE38QQR, INO 1001, IPdR₁ KRX-0402,lucanthone, LY 317615, neuradiab, vitespan, Rta 744, Sdx 102,talampanel, atrasentan, Xr 311, romidepsin, ADS-100380, sunitinib,5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin,liposomal doxorubicin, 5′-deoxy-5-fluorouridine, vincristine,temozolomide, ZK-304709, seliciclib; PD0325901, AZD-6244, capecitabine,L-Glutamic acid,N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-,disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan,tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole,DES(diethylstilbestrol), estradiol, estrogen, conjugated estrogen,bevacizumab, IMC-1C11, CHIR-258,);3-[5-(methylsulfonylpiperadinemethyl)-indolylj-quinolone, vatalanib,AG-013736, AVE-0005, the acetate salt of [D-Ser(Bu t) 6, Azgly 10](pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu t)-Leu-Arg-Pro-Azgly-NH₂ acetate[C₅₉H₈₄N₁₈Oi₄-(C₂H₄O₂)_(x) where x=1 to 2.4], goserelin acetate,leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate,hydroxyprogesterone caproate, megestrol acetate, raloxifene,bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714;TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody,erbitux, EKB-569, PKI-166, GW-572016, Ionafarnib, BMS-214662,tipifarnib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acid,valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951,aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, BacillusCalmette-Guerin (BCG) vaccine, bleomycin, buserelin, busulfan,carboplatin, carmustine, chlorambucil, cisplatin, cladribine,clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin,daunorubicin, diethylstilbestrol, epirubicin, fludarabine,fludrocortisone, fluoxymesterone, flutamide, gemcitabine, hydroxyurea,idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine,mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate,mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin,pamidronate, pentostatin, plicamycin, porfimer, procarbazine,raltitrexed, rituximab, streptozocin, teniposide, testosterone,thalidomide, thioguanine, thiotepa, tretinoin, vindesine,13-cis-retinoic acid, phenylalanine mustard, uracil mustard,estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosinearabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin,mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat,COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668,EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifene,idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab,denileukin diftitox, gefitinib, bortezimib, paclitaxel, cremophor-freepaclitaxel, docetaxel, epithilone B, BMS-247550, BMS-310705,droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA-923, arzoxifene,fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339,ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin,40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001,ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646,wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin,erythropoietin, granulocyte colony-stimulating factor, zolendronate,prednisone, cetuximab, granulocyte macrophage colony-stimulating factor,histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylatedinterferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase,lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane,alemtuzumab, all-transretinoic acid, ketoconazole, interleukin-2,megestrol, immune globulin, nitrogen mustard, methylprednisolone,ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine,bexarotene, tositumomab, arsenic trioxide, cortisone, editronate,mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase,strontium 89, casopitant, netupitant, an NK-1 receptor antagonists,palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide,lorazepam, alprazolam, haloperidol, droperidol, dronabinol,dexamethasone, methylprednisolone, prochlorperazine, granisetron,ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin,epoetin alfa, darbepoetin alfa, and mixtures thereof, among others.

Neuroprotective agents which may be used in combination with compoundsaccording to the present invention include for example, dilostazol,desflurane, eldepryl, felbamate, felbatol, gagitril, latanoprost,methylprednisolone, modafinil, pergolide, permaqx, phentermine andtopiramate (combination), pletal, provigil, osymia, rilutek, riluzole,selegiline, supreme, tiagabine, topamax, topiramate and xalatan, amongothers.

Thrombolytic agents which may be used in combination with compoundsaccording to the present invention include, for example, abbokinase,activase, alteplase, cathflo activase, kinlytic, retavase, reteplase,tenecteplase, tnkase and urokinase, among others.

Immunosuppressant agents which may be used in combination with compoundsaccording to the present invention include, for example, corticosteroids(e.g. prednisolone), ciclosporin (cyclosporine), tacrolimus, andcytotoxic agents (e.g. azathioprine, chlorambucil, cyclophosphamide,methotrexate), among others.

Antioxidants which may be used in combination with compounds accordingto the present invention include, for example, Glutathione, vitamin C,vitamin A, vitamin E, catalase, superoxide dismutase, peroxidases, amongnumerous others.

Other compounds which may be used in combination with compoundsaccording to the present invention include α- or β-adrenergic agonist orantagonist or a NMDA receptor antagonist, all well known in the art.

In some embodiments, the known therapeutic agent(s) is/are administeredaccording to its standard or approved dosing regimen and/or schedule. Insome embodiments, the known therapeutic agent(s) is/are administeredaccording to a regimen that is altered as compared with its standard orapproved dosing regimen and/or schedule. In some embodiments, such analtered regimen differs from the standard or approved dosing regimen inthat one or more unit doses is altered (e.g., reduced or increased) inamount, and/or in that dosing is altered in frequency (e.g., in that oneor more intervals between unit doses is expanded, resulting in lowerfrequency, or is reduced, resulting in higher frequency).

The invention will be more fully understood by reference to thefollowing examples. They should not, however, be construed as limitingthe scope of the invention. All literature citations are incorporated byreference.

EXEMPLIFICATION

The following examples demonstrate, among other things, the CXCR2antagonism and neuroprotective properties of provided peptides (e.g.,ac-PGG) in various embodiments and are not intended to limit the presentclaims.

Example 1 Ac-PGG is an Antagonist of CXCR2 Receptor

This Example shows, among other things, that administration of ac-PGGpeptide results in effects antagonistic to those of effects of ac-PGPmediated by the CXCR2 receptor. These results are particularlysurprising since ac-PGG was previously thought to be an inert controlpeptide with no biological function, and certainly not a potentialtherapeutic action.

First, the mechanism by which ac-PGP induces apoptosis in corticalneurons was investigated. Because ac-PGP is reported to engageneutrophil CXCR2 receptors, whether an established CXCR2 receptorantagonist, SB225002, could influence ac-PGP neurotoxicity was tested.

Primary Cortical Neuron Culture

Cortical neurons were obtained from Sprague-Dawley rat embryos at day 18of gestation. Cortices were triturated digested with 0.025% trypsin inphosphate buffer saline (PBS) for 10 minutes. Digestion was stopped bythe addition of Dulbecco's Modified Eagle Medium (DMEM) containing 10%fetal bovine serum (FBS). Trypsinized cells were filtered using 70 μMdisposable cell filters (Becton-Dickinson), washed twice with Neurobasalmedium (Gibco) containing 4% FBS, and preplated 4 times for 30 minutesin 150 mm vacuum-plasma treated dishes (Becton-Dickinson). During thesepreplating steps, primarily non-neuronal cells adhere to the dish,thereby enriching the neuronal population. Enriched neurons were platedat a density of 50×103/cm2 in Neurobasal medium containing 4% FBS, 25 μMGlutamax (Gibco), and penicillin-streptomycin (Gibco) in whiteclear-bottomed 96-well plates (353377) or 24-well plates (353226)(Becton-Dickinson) coated overnight with 50 μg/ml poly-D-lysine. Atplating, spaces between wells in 96-well plates were filled with 50 μlsterile water to minimize evaporation of medium at the edges of thedish. Twenty-four hours later (1 day in vitro, 1 DIV), 50% of the mediumwas replaced with Neurobasal medium, 2% B27 (Gibco), and 4 μM cytosinearabinoside (AraC) (2 μM AraC final). At 5 DIV, 50% of the medium wasreplaced with Neurobasal, 2% B27. At 7 DIV, cortical cultures were 98%neurons as assessed by microtubule-associated protein 2 (MAP2) antibodylabeling and flow cytometry analysis (data not shown).

Oxygen-Glucose Deprivation (OGD)

Cortical neurons were deprived of oxygen and glucose by incubation indeoxygenated glucose-free Hank's buffered saline solution (HBSS). HBSSwas prepared as follows: sodium chloride, NaCl, 8.0 g/L, potassiumchloride, KCl, 0.4 g/L, potassium phosphate monobasic, KH2PO4, 0.06 g/L,sodium phosphate dibasic, Na2HPO4, 0.048 g/L, magnesium sulfate, MgSO4,0.098 g/L, calcium chloride, CaCl2, 0.14 g/L, sodium bicarbonate,NaHCO3, 0.35 g/L, D-glucose, C6H12O6, 1.0 g/L. Solutions were adjustedto pH 7.4 and sterilized by vacuum filtration through 0.2 μm polyethersulfone (PES) membranes. Glucose-free HBSS was prepared by omittingglucose and adding an additional half molar amount of NaCl to adjustosmolarity (8.162 g/L NaCl). Glucose-free HBSS was deoxygenated in a gaswashing bottle with 95% nitrogen, 5% CO2 for 20 minutes at a flow rateof 3 L/min in a 37° C. water bath. Culture dishes were washed twice withdeoxygenated glucose-free HBSS and incubated for 2 hours in glucose-freedeoxygenated HBSS (OGD). Normoxic control cells were handled identicallybut were incubated in a 5% CO2 incubator with normal HBSSpre-equilibrated in a 5% CO2 atmosphere overnight.

Results

It was found that low nanomolar concentrations of SB225002 blockedac-PGP neurotoxicity as measured via TUNEL stain (FIG. 1 A). Theseresults indicate that ac-PGP-induced apoptosis is CXCR2receptor-dependent in cortical neurons. To further confirm theinvolvement of CXCR2 receptors in ac-PGP mediated neurotoxicity, primaryneurons were incubated with an antibody against the CXCR2 receptorextracellular ligand binding domain prior to ac-PGP exposure. A 1:1000dilution of the antibody blocked ac-PGP-induced apoptosis (FIG. 1 B).Together, these results indicate that ac-PGP initiates neuronalapoptosis through its binding at neuronal CXCR2 receptors. Pretreatmentof neurons with ac-PGG (FIG. 1 C) resulted in significantneuroprotection against ac-PGP exposure similar to that observed withestablished CXCR2 antagonists. These results indicate, for the firsttime, that ac-PGG is a potent CXCR2 antagonist when present inconcentrations as much as 1000 times below the concentration of a CXCR2ligand.

To investigate the possible involvement of ac-PGP and CXCR2 receptors inneuronal death after ischemia/reperfusion, an oxygen-glucose deprivation(OGD) model of stroke in primary neurons was used. Two-hour OGD followedby 24-hour reoxygenation resulted in roughly 40% apoptotic cell death(FIG. 2). Pretreatment with the CXCR2 antagonist SB225002, a CXCR2antibody, or ac-PGG peptide each resulted in significant protection fromOGD. These results indicate a major role for CXCR2 receptor binding inneuronal toxicity after ischemia. Treatment with ac-PGP during OGDresulted in only slightly higher apoptosis than OGD alone. The lack ofan additive effect of OGD and ac-PGP treatment, together with the nearcomplete protection of neurons afforded by CXCR2 blockade, suggests thatCXCR2 receptor binding is a central component of neuronal toxicity afterischemia.

Example 2 PGG is an Antagonist of CXCR2 Receptor

This Example shows, among other things, that administration of ac-PGGpeptide results in effects antagonistic to those of effects of ac-PGPmediated by the CXCR2 receptor.

Primary Cortical Neuron Culture

Cortical neurons were obtained from Sprague-Dawley rat embryos at day 18of gestation. Cortices were triturated digested with 0.025% trypsin inphosphate buffer saline (PBS) for 10 minutes. Digestion was stopped bythe addition of Dulbecco's Modified Eagle Medium (DMEM) containing 10%fetal bovine serum (FBS). Trypsinized cells were filtered using 70 μMdisposable cell filters (Becton-Dickinson), washed twice with Neurobasalmedium (Gibco) containing 4% FBS, and preplated 4 times for 30 minutesin 150 mm vacuum-plasma treated dishes (Becton-Dickinson). During thesere Latin steps, primarily non-neuronal cells adhere to the dish, therebyenriching the neuronal population. Enriched neurons were plated at adensity of 50×103/cm2 in Neurobasal medium containing 4% FBS, 25 μMGlutamax (Gibco), and penicillin-streptomycin (Gibco) in whiteclear-bottomed 96-well plates (353377) or 24-well plates (353226)(Becton-Dickinson) coated overnight with 50 μg/ml poly-D-lysine. Atplating, spaces between wells in 96-well plates were filled with 50 μlsterile water to minimize evaporation of medium at the edges of thedish. Twenty-four hours later (1 day in vitro, 1 DIV), 50% of the mediumwas replaced with Neurobasal medium, 2% B27 (Gibco), and 4 μM cytosinearabinoside (AraC) (2 μM AraC final). At 5 DIV, 50% of the medium wasreplaced with Neurobasal, 2% B27. At 7 DIV, cortical cultures were 98%neurons as assessed by microtubule-associated protein 2 (MAP2) antibodylabeling and flow cytometry analysis (data not shown).

Oxygen-Glucose Deprivation (OGD)

Cortical neurons were deprived of oxygen and glucose by incubation indeoxygenated glucose-free Hank's buffered saline solution (HBSS). HBSSwas prepared as follows: sodium chloride, NaCl, 8.0 g/L, potassiumchloride, KCl, 0.4 g/L, potassium phosphate monobasic, KH2PO4, 0.06 g/L,sodium phosphate dibasic, Na2HPO4, 0.048 g/L, magnesium sulfate, MgSO4,0.098 g/L, calcium chloride, CaCl2, 0.14 g/L, sodium bicarbonate,NaHCO3, 0.35 g/L, D-glucose, C6H12O6, 1.0 g/L. Solutions were adjustedto pH 7.4 and sterilized by vacuum filtration through 0.2 μm polyethersulfone (PES) membranes. Glucose-free HBSS was prepared by omittingglucose and adding an additional half molar amount of NaCl to adjustosmolarity (8.162 g/L NaCl). Glucose-free HBSS was deoxygenated in a gaswashing bottle with 95% nitrogen, 5% CO2 for 20 minutes at a flow rateof 3 L/min in a 37° C. water bath. Culture dishes were washed twice withdeoxygenated glucose-free HBSS and incubated for 2 hours in glucose-freedeoxygenated HBSS (OGD). Normoxic control cells were handled identicallybut were incubated in a 5% CO2 incubator with normal HBSSpre-equilibrated in a 5% CO2 atmosphere overnight.

Results

It was found that pretreatment of neurons with PGG (FIG. 3) resulted insignificant neuroprotection against ac-PGP exposure similar to thatobserved with established CXCR2 antagonists. These results indicate,that PGG is a potent CXCR2 antagonist when present in concentrations asmuch as 1000 times below the concentration of a CXCR2 ligand.

Example 2 Neuroprotective Effect of Ac-PGG on Ac-PGP-Induced Apoptosis

This Example shows, among other things, that administration of an ac-PGGpeptide results in neuroprotective effects via the CXCR2 receptor.

The efficacy of CXCR2 antagonist to prevent neuronal death when given atreoxygenation or at time points up to an hour after reoxygenation wastested (FIG. 4A-C). Primary cortical neuron culture and oxygen-glucosedeprivation (OGD) methods, as described above in Example 1, were used toproduce the results in this example. As shown in FIG. 4A-C, each ofSB2225002, a CXCR2 antibody, and ac-PGG provided neuroprotection whengiven at reoxygenation and at later time points up to 60 minutespost-reoxygenation. However, the effectiveness of SB225002 significantlydeclined at 15 minutes post-reoxygenation (FIG. 4A) and the CXCR2antibody was significantly less protective at 60 minutespost-reoxygenation (FIG. 4B). Ac-PGG treatment was equally as effectiveat each time point measured (FIG. 4C). These results indicate thatac-PGG is an effective CXCR2 antagonist and neuroprotectant throughoutan extended time window after ischemia.

Example 3 Neuroprotective Effect of Ac-PGG in Model of Stroke

This Example shows, among other things, that administration of an ac-PGGpeptide results in neuroprotective effects in a model of stroke.

Ac-PGG Formulation and Administration

At reperfusion, animals received 0.5 mL of normal saline vehicle or 0.3,3, or 30 mg ac-PGG (1, 10, or 100 mg/kg, respectively) in 0.5 mL normalsaline by IV injection through the lateral tail vein.

Stroke Model with Spontaneously Hypertensive Rats (SHR)

Male spontaneously hypertensive rats (SHR) weighing 290-300 g wereobtained from Charles River Laboratories. Animals were anesthetized with2% isoflurane inhalant during all surgical procedures. The right commoncarotid artery (CCA) was ligated approximately 6 mm caudal to thebifurcation of the external and internal carotid arteries. With avascular clamp in place on the CCA immediately caudal to thebifurcation, an incision was made in the CCA with a 25 G needle 5 mmcaudal to the clamp and a 4-0 nylon suture (Doccol Corporation) 30 mm inlength with a 2-3 mm silicone coated tip (0.39 mm diameter) was advancedinto the CCA lumen and secured in place with 2 4-0 silk sutures spaced 3mm apart and tied around the CCA caudal to the clamp. The clamp wasremoved and the suture advanced into the internal carotid artery untilresistance was felt. In this position, the filament occluded the middlecerebral artery (MCA) origin and MCA territory became ischemic. Thesuture was secured with the 2 silk sutures around the CCA. The wound wasclosed and the animal was allowed to recover during the 90-minuteocclusion period. After 90 minutes, the suture was removed and the CCAligated rostral to the incision.

All animals received 3 mL of subcutaneous saline after surgery toprevent dehydration. After 24 hours reperfusion, brains were sectionedinto 4 mm-thick quadrants, and infarcted tissue identified by2,3,5-triphenyl-tetrazolium chloride (TTC) staining. Edema-correctedinfarct volume was calculated by subtracting the area of non-infarctedtissue in the ipsilateral hemisphere from the total volume of thecontralateral hemisphere. Infarct volume was quantified using Image Jsoftware.

Results

As shown in FIG. 5, treatment with ac-PGG at the time of reperfusionsignificantly decreased total infarct volume compared to treatment withVehicle. Additionally, as shown in FIG. 6, treatment with ac-PGG at thetime of reperfusion significantly increased the percent reduction ininfarct volume. FIG. 7, shows representative TTC-stained brain sectionsdemonstrating differences in infarction between animals receivingvehicle or 100 mg/kg ac-PGG. These results indicate that treatment withac-PGG can be effective in a stroke model after ischemia, which hasclinical relevance due to the real-world necessity to administertreatment hours to days after a stroke.

Example 4 Anti-Inflammatory Effect of Ac-PGG on Carrageenan-Induced PawEdema

This Example shows, among other things, that administration of ac-PGGpeptide results in decreased paw volume and increased latency toresponse in hot plate test.

Ac-PGG Formulation

Ac-PGG dose volume was 2 mL/kg for IV injections. Ac-PGG was freshlydissolved in saline to a concentration of 200 mg/mL (stock solution) andthe working solution of Ac-PGG was diluted in saline at a concentrationof 20 mg/mL, 2 mg/mL and 0.2 mg/mL for dose levels of 100 mg/kg, 10mg/kg, 1 mg/kg and 0.1 mg/kg, respectively.

Carrageenan-Induced Edema

On Day 1, a 1% WN solution of λ-Carrageenan in saline (0.9% NaCl) wasprepared. All animals were lightly anesthetized with isoflurane. Abaseline measurement of left hind paw volume for each animal wasperformed before Carrageenan injection occurred. 100 μL of 1%Carrageenan solution was injected subcutaneously (s.c.) into the planterpad of the left hind paw using a microsyringe.

Paw Volume Measurements

Volumes of the paw injected with Carrageenan were measured with calipersjust prior to Carrageenan administration (time 0) and then measuredagain 1, 3, 4, and 5 hours after Carrageenan injection. Paws weremeasured in two axes and then paw volume was calculated.

Paw edema volume in this model is an indication of inflammationseverity. For each time point, change in paw volume was calculatedeither by subtracting the baseline paw volume or by determining the % ofbaseline paw volume.

Hot Plate Test Measurements

One hour before and 3 and 5 hours post-Carrageenan injection, the ratswere transported to a testing room and left undisturbed for at least 15minutes before the test. Illumination in the testing room was 100-130Lux at the level of the bench (red light). The hot plate was maintainedthermostatically at a temperature of 50° C. One rat at a time was placedon the hot plate platform and latency to jumping or lifting theCarrageenan-injected paw or the control paw was recorded. The latency ofthe response was measured in a blinded manner. In the absence of aresponse, a 30 second cut-off was used to prevent tissue damage.

Ac-PGG Administration

Ac-PGG was administered by a single intravenous (IV) injection half anhour before Carrageenan injection. See Table 1 for group allocations.

TABLE 1 Group Treatment Dose Administration Total rats 1 Vehicle 0 IV 72 Ac-PGG 0.1 mg/kg  IV 7 3 Ac-PGG 1 mg/kg IV 7 4 Ac-PGG 10 mg/kg  IV 7 5Ac-PGG 100 mg/kg  IV 7 6 Indomethacin 2 mg/kg IP 7

Results

The 3 highest doses of ac-PGG decreased the Carrageenan-induced increasein paw volume. As shown in FIG. 8, the 3 highest doses of ac-PGGdecreased the Carrageenan-induced increase in paw volume at 3, 4, and 5hours post-Carrageenan injection. Additionally, the 50 mg/kg dose ofac-PGG significantly decreased the change in paw volume compared to theVehicle group after 1 hour. Also, even the 0.1 mg/kg dose of ac-PGGsignificantly decreased the change in paw volume 5 hourspost-Carrageenan injection.

Treatment with ac-PGG significantly increased latency to respond to thehot plate. As shown in FIG. 9, the 1 mg/kg and 50 mg/kg doses of ac-PGGdecreased change in latency to hot plate response 3 hours afterCarrageenan injection, while only the 50 mg/kg dose lead to asignificant decrease in latency change 5 hours after Carrageenaninjection.

Example 5 Anti-Inflammatory Effect of Ac-PGG on EAE-Induced Paralysis

This Example shows, among other things, that administration of ac-PGGpeptide results in decreased paralysis and weight loss in anexperimental autoimmune encephalomyelitis (EAE) model.

Ac-PGG Formulation

Ac-PGG dose was 100 mg/kg for subcutaneous injections. Treatment animalsreceived 30 mg ac-PGG in 200 μL normal saline.

EAE Induction

Male Lewis rats (10 weeks old) were obtained from Charles RiverLaboratories. After a 1-week acclimation period, EAE was induced byinjecting animals subcutaneously at the base of the tail with 100 μgguinea pig myelin basic protein in 50 μL normal Saline mixed with 150 μLFreund's complete adjuvant for a 200 μL total injection volume.

EAE Scoring

Animals were weighed and assigned an EAE score daily for 25 days. Thecriteria used for EAE scoring are presented in Table 2.

TABLE 2 Score Sign 0 Normal 1 Weak distal tail 2 Weak proximal tail 3Tail paralysis 4 Tail paralysis and hindlimb weakness 5 Partial hindlimbparalysis (one limb) 6 Hindlimb paralysis 7 Hindlimb paralysis andforelimb weakness 8 Hindlimb paralysis, forelimb weakness, and sideresting position 9 Hindlimb paralysis, forelimb paralysis, and sideresting position 10 Hindlimb paralysis, forelimb paralysis, andunresponsive to stimuli

Ac-PGG Administration

Ac-PGG was administered by subcutaneous injection on the hindquartersstarting with the left hindquarter and alternating sides with eachinjection. Treatment animals received ac-PGG on days 0-7 or 8-15 andreceived 200 μL subcutaneous injections of saline on days when ac-PGGwas not administered. Vehicle animals were administered 200 μL normalsaline on days 0-15 by subcutaneous injection on the hindquartersstarting with the left hindquarter and alternating sides with eachinjection.

Results

Both ac-PGG dosing schedules (days 0-7 and days 8-15) decreased theEAE-induced paralysis. As shown in FIG. 10, both ac-PGG dosing schedulessignificantly decreased both EAE scores pooled over 26 days and Day 14EAE scores. Additionally, as shown in FIG. 11, both ac-PGG dosingschedules significantly increased weight compared to the Vehicle groupover the course of the study and there was a significant differencebetween the final weights of the Vehicle group and the ac-PGG (days8-15) group on Day 25.

Ac-PGG also decreased white matter hyperintensities as detected byT2-weighted MRI. As shown in FIG. 12, in animals treated with ac-PGG ondays 8-15, fewer coronal brain sections contained white matterhyperintensities on Day 14 of EAE induction compared to Vehicle-treatedanimals.

EQUIVALENTS AND SCOPE

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. The scope of the presentinvention is not intended to be limited to the above Description, butrather is as set forth in the following claims:

1-86. (canceled)
 87. A pharmaceutical composition comprising aneffective amount of at least one compound selected from the groupconsisting of a tripeptide with the amino acid sequenceproline-glycine-glycine (PGG tripeptide), a PGG tripeptide which isacetylated or methylated at its N-terminus, a PGG tripeptide which isamidated at its C-terminus, a PGG tripeptide which is PEGylated, apharmaceutically acceptable salt thereof, and mixtures thereof,optionally in combination with a pharmaceutically acceptable carrier orexcipient.
 88. The composition according to claim 87, wherein saidcompound is the tripeptide with the amino acid sequenceproline-glycine-glycine (PGG), a pharmaceutically acceptable saltthereof or a mixture thereof.
 89. The composition according to claim 87wherein said compound is the tripeptide which is acetylated ormethylated at the N-terminus, a pharmaceutically acceptable salt thereofor a mixture thereof.
 90. The composition according to claim 89 whereinsaid compound is the tripeptide which is acetylated at the N-terminus.91. The composition according to claim 87 wherein said compound is thetripeptide which is amidated at the C-terminus.
 92. The compositionaccording to claim 87 wherein said compound is the tripeptide which isPEGylated.
 93. A pharmaceutical composition comprising a tripeptide withthe amino acid sequence proline-glycine-glycine (PGG) wherein thepeptide is acetylated or methylated at the N-terminus, PEGylated, oramidated at the C-terminus in any combination thereof.
 94. Thepharmaceutical composition of claim 87 which is adapted for intravenous,subcutaneous, oral, transdermal, aerosol inhalation, intramuscular, orintraperitoneal administration.
 95. The pharmaceutical composition ofclaim 93 which is adapted for intravenous, subcutaneous, oral,transdermal, aerosol inhalation, intramuscular, or intraperitonealadministration.
 96. The pharmaceutical composition according to claim 87further including an effective amount of at least one anti-inflammatoryagent, anticancer agent, neuroprotective agent, thrombolytic agent,immunosuppressant, antioxidant, α- or β-adrenergic agonist orantagonist, or NMDA receptor antagonist.
 97. The pharmaceuticalcomposition according to claim 93 further including an effective amountof at least one anti-inflammatory agent, anticancer agent,neuroprotective agent, thrombolytic agent, immunosuppressant,antioxidant, α- or β-adrenergic agonist or antagonist, or NMDA receptorantagonist.
 98. A method of treating inflammation in a patient orsubject in need comprising administering to said patient or subject aneffective amount of a composition according to claim
 87. 99. A method oftreating inflammation in a patient or subject in need comprisingadministering to said patient or subject an effective amount of acomposition according to claim
 93. 100. A method of treatinginflammation in a patient or subject in need comprising administering tosaid patient or subject an effective amount of a composition accordingto claim
 96. 101. A method of treating inflammation in a patient orsubject in need comprising administering to said patient or subject aneffective amount of a composition according to claim
 97. 102. The methodof claim 98, wherein the inflammation is associated with inflammatoryconditions of the nervous system (e.g., stroke, traumatic brain injury,Alzheimer's disease, amyotrophic lateral sclerosis (ALS), multiplesclerosis, Parkinson's disease, neuropathy, neuropathic pain), liver(e.g., hepatitis), kidney (e.g. nephritis), gastrointestinal tract(e.g., inflammatory bowel disease, Crohn's disease, ulcerative colitis),and pancreas (e.g., diabetes, pancreatitis); cardiovascular disorders(e.g., pulmonary fibrosis, reperfusion injury, vasculitis,vaso-occlusive crisis) respiratory disorders (e.g., cystic fibrosis,chronic obstructive pulmonary disease); arthritis (e.g., rheumatoidarthritis); autoimmune disorders (e.g., thyroiditis, scleroderma);traumatic injury; infectious disease (e.g., acute endocarditis,pericarditis); tumors or cancers (e.g., soft tissue or solid tumors);transplant rejection (e.g., destruction of pancreatic islet cells inislet cell transplantation, delayed graft failure in kidney or otherorgan transplantation); acute and chronic graft versus host disease.103. The method of claim 98, wherein said inflammation is associatedwith amyotrophic lateral sclerosis, Huntington's disease, Alzheimer'sdisease, Parkinson's disease, multiple sclerosis, vascular cognitiveimpairment, stroke, traumatic brain injury, spinal cord injury,neuropathy, encephalitis, epilepsy, prion diseases, and combinationthereof.
 104. The method of claim 98, wherein said inflammation isassociated with stroke, multiple sclerosis (MS), amyotrophic lateralsclerosis (ALS), Alzheimer's disease or Parkinson's disease.
 105. Themethod of claim 99, wherein the inflammation is associated withinflammatory conditions of the nervous system (e.g., stroke, traumaticbrain injury, Alzheimer's disease, amyotrophic lateral sclerosis (ALS),multiple sclerosis, Parkinson's disease, neuropathy, neuropathic pain),liver (e.g., hepatitis), kidney (e.g. nephritis), gastrointestinal tract(e.g., inflammatory bowel disease, Crohn's disease, ulcerative colitis),and pancreas (e.g., diabetes, pancreatitis); cardiovascular disorders(e.g., pulmonary fibrosis, reperfusion injury, vasculitis,vaso-occlusive crisis) respiratory disorders (e.g., cystic fibrosis,chronic obstructive pulmonary disease); arthritis (e.g., rheumatoidarthritis); autoimmune disorders (e.g., thyroiditis, scleroderma);traumatic injury; infectious disease (e.g., acute endocarditis,pericarditis); tumors or cancers (e.g., soft tissue or solid tumors);transplant rejection (e.g., destruction of pancreatic islet cells inislet cell transplantation, delayed graft failure in kidney or otherorgan transplantation); acute and chronic graft versus host disease.106. The method of claim 99, wherein said inflammation is associatedwith amyotrophic lateral sclerosis, Huntington's disease, Alzheimer'sdisease, Parkinson's disease, multiple sclerosis, vascular cognitiveimpairment, stroke, traumatic brain injury, spinal cord injury,neuropathy, encephalitis, epilepsy, prion diseases, and combinationthereof.
 107. The method of claim 99, wherein said inflammation isassociated with stroke, multiple sclerosis (MS), amyotrophic lateralsclerosis (ALS), Alzheimer's disease or Parkinson's disease.